双吲哚马来酰亚胺衍生物及其制备方法和用途
技术领域:
本发明涉及双吲哚马来酰亚胺衍生物及其制备方法和用途。
背景技术:
随着人们生活习惯和膳食结构的变化,糖尿病的发病率逐年上升。据2011年WHO发布的数据显示,每年全世界约有460万人死于糖尿病相关的疾病,用于糖尿病的医疗费用高达4650亿美元。目前,中国有近1.0亿名糖尿病患者以及约3.4亿糖代谢异常人群。糖尿病已经成为继癌症、心血管疾病之后的第三大杀手。
糖尿病分为:1型糖尿病(胰岛素依赖型糖尿病)和2型糖尿病(非胰岛素依赖型糖尿病),超过90%的病人属于2型糖尿病。2型糖尿病患者无需胰岛素治疗,可饮食调控血糖,仅在饮食调节无效时用药物辅助治疗。食物中的淀粉经口腔唾液、胰淀粉酶消化成葡萄糖分子的低聚糖,α-葡萄糖苷酶的作用是在非还原末端断裂这些低聚糖的α-1,4-糖苷键,释放出葡萄糖,葡萄糖被小肠上皮吸收后进入血液循环,成为血糖。在生理状态下,小肠上、中、下三段均存在α-葡萄糖苷酶,因此人体对糖的吸收迅速而完善。α-葡萄糖苷酶抑制剂通过可逆或竞争性抑制小肠刷状缘的α-葡萄糖苷酶的活性,延迟或减缓餐后血糖的升高。此外,空腹(低位小肠已无食物成分时)服用α-葡萄糖苷酶抑制剂类药物后,肠内碳水化合物、脂肪、蛋白质等食糜进入回肠远端;该部位是小肠胰升糖素样肽21(GLP21)储量最丰富的位置,可以刺激GLP21分泌的增加,刺激胰岛素的释放,从而降低餐后血糖浓度。因此,α-葡萄糖苷酶抑制剂(AGIs)是一类以延缓肠道糖类吸收而达到治疗糖尿病的口服降糖药物。
目前,临床上应用的α-葡萄糖苷酶抑制剂类药物主要是:阿卡波糖、伏格列波糖和米格列醇,服用这三类药后,由于本身是不分解糖类物质而直接到达大肠,会出现腹部不适、胀气、排气等不良反应。故需寻找新的α-葡萄糖苷酶抑制剂类药物。
糖尿病肾病(Diabetic Kidney Disease,DKD)是糖尿病最常见的严重微血管并发症之一,发生率约为糖尿病患者的30%-40%,已成为终末期肾病(End Stage RenalDisease,ESRD)的第一位原因,也是糖尿病致命的主要原因。目前虽然在抗DKD药物研究方面已取得一定的进展,但是现有的临床治疗如控制血压、血糖、血脂以及抗炎等手段只能延缓DKD向肾功能衰竭的进程,而无法逆转或阻止病情发展;同时也面临疗效不肯定、毒副作用大等问题(Curr Diabetes Rev 2005;1:281-286),因此迫切需要研发针对新作用靶点且安全高效的新型抗DKD药物。
发明内容:
双吲哚马来酰亚胺衍生物因其结构多样性和良好的生物活性受到广泛关注,本发明人致力于双吲哚马来酰亚胺衍生物的药物开发,意外地发现,该类化合物具有α-葡萄糖苷酶抑制活性和抗糖尿病作用。
由此,本发明人提供一种式A化合物、其药学上可接受的盐或前药:
其中,
虚线表示没有化学键或为单键;
R1和R2各自独立地选自:-H;烷基,所述烷基任选被氨基、氰基、羟基、羧基、烷氧基、脂杂环基、芳基、杂芳基、-COA取代;烯基,所述烯基任选被氨基、氰基、羟基、羧基、烷氧基、脂杂环基、芳基、杂芳基、-COA取代;单糖基,所述单糖基的羟基氢任选被烷基取代;其中A选自氢,-NR13R14,芳基,芳氨基,任选被羟基、卤素取代的烷基,任选被羟基、卤素取代的烷氧基;
或者,R1和R2一起构成-(CH2)m1-O-(CH2)m2-,其中的H任选被-(CH2)0~8-NR13R14取代,m1和m2各自独立地为1~6的整数;
或者,R1和R2一起构成如下基团:
其中,R9、R10独立地为-H或烷基;或者,R9与R10一起构成-C(=O)-;
R8选自:-H;羟基;烷基,所述烷基任选被烷氧基取代;烯基;炔基;芳基,所述芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;脂杂环基,所述脂杂环基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;杂芳基,所述杂芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;-C(=Y1)-Y2;-S(=O)2-Y3;
Y1选自:=O;=S;=NH;
Y2选自:烷基;烷氧基;羟胺基;-NR13R14;芳基,所述芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;杂芳基,所述杂芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;脂杂环基,所述的脂杂环基均任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;脂杂环基取代的烷基取代的脂杂环基,所述的脂杂环基均任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;烷氨基,所述烷氨基被氧、羟基、杂芳基、芳基中至少一种取代,所述杂芳基、芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;
Y3为任选被卤素、卤代烷基取代的芳基;
R3选自-H;羟基;卤素;-NR13R14;-(C=O)NR11R12;-(C=O)R15;-O(C=O)R16;-NR17-(C=O)R18;-NR19-(COO)R20;-NR21-(SO2)R22;-O(C=O)NR23R24;-SR25;-(S=O)R26;-(SO2)R27;-(SO2)NR32R33;烷基,所述烷基任选被羟基、氰基、羧基、单糖基、烷氧基、芳基、杂芳基、脂杂环基、-NH-CO-亚烷基(NH2)(A1)、-NR13R14取代,所述芳基、杂芳基、脂杂环基任选被氨基、羟基、卤素、烷基、卤代烷基取代;烯基,所述烯基任选被羟基、氰基、羧基、单糖基、烷氧基、芳基、杂芳基、脂杂环基、-NH-CO-亚烷基(NH2)(A1)、-NR13R14取代;炔基,所述炔基任选被羟基、氰基、羧基、单糖基、烷氧基、芳基、杂芳基、脂杂环基、-NH-CO-亚烷基(NH2)(A1)、-NR13R14取代;
A1选自-H、任选被杂芳基取代的烷基;
R4与R5一起构成=O,和/或R6与R7一起构成=O;
当R4与R5或R6与R7不构成=O时,各自独立地选自-H;羟基;-NR13R14;-(C=O)R15;-NR17-(C=O)R18;-SR25;芳基,所述芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;杂芳基,所述杂芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;烷氧基,所述烷氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;芳氧基,所述芳氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;
G1~G8各自独立地选自-H;卤素;羟基;氰基;硝基;羧基;-NR13R14;-(C=O)NR11R12;-(C=O)R15;-O(C=O)R16;-NR17-(C=O)R18;-NR19-(COO)R20;-NR21-(SO2)R22;-O(C=O)NR23R24;-SR25;-(S=O)R26;-(SO2)R27;-CH=NOR28;-CH=NR29;-CH=NNR30R31;-(SO2)NR32R33;烷基,所述烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0- 2R34取代;烯基,所述烯基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;炔基,所述炔基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;芳基,所述芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;杂芳基,所述杂芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;烷氧基,所述烷氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;芳氧基,所述芳氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;
R13、R14各自独立地选自-H;氨基;单糖基,所述单糖基的羟基氢任选被烷基取代;烷基,所述烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;芳基,所述芳基任选被氨基、羟基、卤素、烷基取代,所述烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;脂杂环基,所述脂杂环基任选被氨基、羟基、卤素、烷基取代,所述烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;
R11、R12、R15、R16、R17、R18、R19、R20、R21、R22、R23、R24、R25、R26、R27、R28、R29、R30、R31、R32、R33、R34各自独立地选自:-H;烷基;芳基;
所述脂杂环基和杂芳基各自独立地含有1-4个杂原子,所述杂原子选自N、O、S。
可选地,上述式A化合物、其药学上可接受的盐或前药中:所述烷基和卤代烷基、烷氧基、烷氨基中的烷基为C1~C20烷基,或者为C1~C18烷基,或者为C1~C6烷基,或者为C1~C4烷基;所述烯基为C2~C20烯基,或者为C2~C18烯基,或者为C2~C6烯基,或者为C2~C4烯基;所述炔基为C2~C20炔基,或者为C2~C18炔基,或者为C2~C6炔基,或者为C2~C4炔基;所述脂杂环基为4~14元单环或多环脂杂环基,环上杂原子数为1~3,或者环上杂原子数为1~2;所述芳基和芳氧基中的芳基为C6~C14单环或多环芳基;所述杂芳基为5~14元单环或多环杂芳基,环上杂原子数为1~3,或者环上杂原子数为1~2。
可选地,上述式A化合物、其药学上可接受的盐或前药中:
R1和R2各自独立地选自:-H;C1~C6烷基,所述C1~C6烷基任选被氨基、氰基、羟基、羧基、C1~C6烷氧基、5或6元脂杂环基、C6~C10芳基、5~10元杂芳基、-COA取代;C2~C6烯基,所述C2~C6烯基任选被氨基、氰基、羟基、羧基、C1~C6烷氧基、5或6元脂杂环基、C6~C10芳基、5~10元杂芳基、-COA取代;单糖基,所述单糖基的羟基氢任选被C1~C6烷基取代;其中A选自氢,-NR13R14,C6~C10芳基,C6~C10芳氨基,任选被羟基、卤素取代的C1~C6烷基,任选被羟基、卤素取代的C1~C6烷氧基;
或者,R1和R2一起构成如下基团:
其中,R9、R10独立地为-H或C1~C6烷基;或者,R9与R10一起构成-C(=O)-;
R8选自:-H;羟基;C1~C18烷基,所述C1~C18烷基任选被C1~C6烷氧基取代;C2~C18烯基;C2~C18炔基;C6~C10芳基,所述C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5或6元脂杂环基,所述脂杂环基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5~10元杂芳基,所述杂芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;-C(=Y1)-Y2;-S(=O)2-Y3;
Y1选自:=O;=S;=NH;
Y2选自:C1~C18烷基;C1~C18烷氧基;羟胺基;-NR13R14;C6~C10芳基,所述C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5~10元杂芳基,所述杂芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5或6元脂杂环基,所述的脂杂环基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5或6元脂杂环基取代的C1~C6烷基取代的5或6元脂杂环基,所述的脂杂环基均任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;C1~C6烷氨基,所述C1~C6烷氨基被氧、羟基、5~10元杂芳基、C6~C10芳基中至少一种取代,所述5~10元杂芳基、C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;
Y3为任选被卤素、卤代C1~C6烷基取代的C6~C10芳基;
R3选自-H;羟基;卤素;-NR13R14;-(C=O)NR11R12;-(C=O)R15;-O(C=O)R16;-NR17-(C=O)R18;-NR19-(COO)R20;-NR21-(SO2)R22;-O(C=O)NR23R24;-SR25;-(S=O)R26;-(SO2)R27;-(SO2)NR32R33;C1~C6烷基,所述C1~C6烷基任选被羟基、氰基、羧基、单糖基、C1~C6烷氧基、C6~C10芳基、5~10元杂芳基、5或6元脂杂环基、-NH-CO-C1~C6亚烷基(NH2)(A1)、-NR13R14取代,所述C6~C10芳基、5~10元杂芳基、5或6元脂杂环基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;C2~C6烯基,所述C2~C6烯基任选被羟基、氰基、羧基、单糖基、C1~C6烷氧基、C6~C10芳基、5~10元杂芳基、5或6元脂杂环基、-NH-CO-C1~C6亚烷基(NH2)(A1)、-NR13R14取代;C2~C6炔基,所述C2~C6炔基任选被羟基、氰基、羧基、单糖基、C1~C6烷氧基、C6~C10芳基、5~10元杂芳基、5或6元脂杂环基、-NH-CO-C1~C6亚烷基(NH2)(A1)、-NR13R14取代;
A1选自-H、任选被5~10元杂芳基取代的C1~C6烷基;
R4与R5一起构成=O,和/或R6与R7一起构成=O;
当R4与R5或R6与R7不构成=O时,各自独立地选自-H;羟基;-NR13R14;-(C=O)R15;-NR17-(C=O)R18;-SR25;C6~C10芳基,所述C6~C10芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;5~10元杂芳基,所述杂芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C1~C6烷氧基,所述C1~C6烷氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C6~C10芳氧基,所述C6~C10芳氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;
G1~G8各自独立地选自-H;卤素;羟基;氰基;硝基;羧基;-NR13R14;-(C=O)NR11R12;-(C=O)R15;-O(C=O)R16;-NR17-(C=O)R18;-NR19-(COO)R20;-NR21-(SO2)R22;-O(C=O)NR23R24;-SR25;-(S=O)R26;-(SO2)R27;-CH=NOR28;-CH=NR29;-CH=NNR30R31;-(SO2)NR32R33;C1~C6烷基,所述C1~C6烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C2~C6烯基,所述C2~C6烯基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C2~C6炔基,所述C2~C6炔基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C6~C10芳基,所述C6~C10芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;5~10元杂芳基,所述杂芳基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C1~C6烷氧基,所述C1~C6烷氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C6~C10芳氧基,所述C6~C10芳氧基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;
R13、R14各自独立地选自-H;氨基;单糖基,所述单糖基的羟基氢任选被C1~C6烷基取代;C1~C6烷基,所述C1~C6烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;C6~C10芳基,所述C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷基取代,所述C1~C6烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;5或6元脂杂环基,所述脂杂环基任选被氨基、羟基、卤素、C1~C6烷基取代,所述C1~C6烷基任选被选自卤素,羟基,氰基,硝基,羧基,叠氮基,-NR13R14,-S(O)0-2R34取代;
R11、R12、R15、R16、R17、R18、R19、R20、R21、R22、R23、R24、R25、R26、R27、R28、R29、R30、R31、R32、R33、R34各自独立地选自:-H;C1~C6烷基;C6~C10芳基;
所述脂杂环基和杂芳基各自独立地含有1-4个杂原子,所述杂原子选自N、O、S。
可选地,上述式A化合物、其药学上可接受的盐或前药为:下述式I化合物、式II化合物或它们药学上可接受的盐或前药,
其中,式I中,
虚线表示没有化学键或为单键;
G1~G8各自独立地选自-H;卤素;C1~C6的烷基;C2~C6烯基;C2~C6炔基;
R3选自-H;-NR13R14;C1~C6烷基,所述C1~C6烷基任选被羟基、氰基、羧基、单糖基、C1~C6烷氧基、C6~C10芳基、5~10元杂芳基、5或6元脂杂环基、-NH-CO-C1~C6亚烷基(NH2)(A1)、-NR13R14取代,所述5或6元脂杂环基、5~10元杂芳基、C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;
其中A1选自:-H;C1~C6烷基,所述C1~C6烷基任选被5~10元杂芳基取代,所述5~10元杂芳基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;R13、R14各自独立地选自:-H;C1~C6烷基;
R1和R2各自独立地选自:-H;C1~C6烷基,所述C1~C6烷基任选被氰基、羟基、羧基、C6~C10芳基、5~10元杂芳基取代,所述C6~C10芳基、5~10元杂芳基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;
式II中,
G1~G8各自独立地选自-H;卤素;
R3选自-H;C1~C6烷基,所述C1~C6烷基任选被羟基、氰基、羧基、单糖基、C1~C6烷氧基、C6~C10芳基、5~10元杂芳基、5或6元脂杂环基、-NH-CO-C1~C6亚烷基(NH2)(A1)、-NR13R14取代,所述5或6元脂杂环基、5~10元杂芳基、C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;
其中A1选自:-H;C1~C6烷基,所述C1~C6烷基任选被5~10元杂芳基取代,所述5~10元杂芳基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;R13、R14各自独立地选自:-H;C1~C6烷基;
R4与R5一起构成=O,和/或R6与R7一起构成=O;
当R4与R5或R6与R7不构成=O时,各自独立地选自-H或-OR35,R35为-H或者C1~C6烷基;
R9、R10独立地为-H或C1~C6烷基;R8选自-C(=Y1)-Y2,-S(=O)2-Y3;
其中,Y1选自=O;=S;=NH;
Y2选自羟胺基;C6~C10芳基,所述C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5~10元杂芳基,所述5~10元杂芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5或6元脂杂环基,所述5或6元脂杂环基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;5或6元脂杂环基取代的C1~C6烷基取代的5或6元脂杂环基,所述5或6元脂杂环基均任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;C1~C6烷氨基,所述C1~C6烷氨基被氧、羟基、5~10元杂芳基、C6~C10芳基中至少一种取代,所述5~10元杂芳基、C6~C10芳基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;
Y3为任选被卤素、卤代C1~C6烷基取代的C6~C10芳基;
或者,R9与R10一起构成-C(=O)-,R8选自-H;C1~C18烷基;C1~C6烷氧基取代的C1~C18烷基。
上述式A化合物、其药学上可接受的盐或前药中:
可选地,式I中,
虚线表示没有化学键或为单键;
G1~G8如权利要求4中所述式I化合物中的定义;
R3选自-H;-NR13R14;C1~C6烷基,所述C1~C6烷基任选被羟基、氰基、羧基、单糖基、C1~C6烷氧基、吗啉基、哌啶基、哌嗪基、吡啶基、苯基或-NH-CO-C1~C6亚烷基(NH2)(A1)、-NR13R14取代,所述吗啉基、哌啶基、哌嗪基、吡啶基、苯基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;
其中A1选自:-H;C1~C6烷基,所述C1~C6烷基任选被咪唑基、吲哚基取代,所述咪唑基、吲哚基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;R13、R14各自独立地选自:-H;C1~C6烷基;
R1和R2各自独立地选自-H;C1~C6烷基,所述C1~C6烷基任选被氰基、羟基、羧基、苯基、萘基、吡啶基取代,所述苯基、萘基、吡啶基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;
可选地,式II中,
G1~G8如权利要求4中所述式II化合物中的定义;
R3选自-H;C1~C6烷基,所述C1~C6烷基任选被氨基、羟基、苯基、吗啉基、哌啶基、哌嗪基取代,所述苯基、吗啉基、哌啶基、哌嗪基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;
R4与R5一起构成=O,和/或R6与R7一起构成=O;
当R4与R5或R6与R7不构成=O时,各自独立地选自-H或-OH;
R9、R10为甲基;
R8选自-C(=Y1)-Y2;-S(=O)2-Y3;
其中,Y1选自=O;=S;=NH;
Y2选自:羟胺基;苯基,所述苯基任选被氨基、羟基、卤素、C1~C6烷氧基、C1~C6烷基、C1~C6卤代烷基取代;咪唑基;哌嗪基,所述哌嗪基任选被吗啉基取代;烷氨基,所述烷氨基被氧、羟基、吲哚基、苯基中至少一种取代;
Y3为任选被卤素、C1~C6卤代烷基取代的苯基;
或者,R9与R10一起构成-C(=O)-,R8选自-H;C1~C18烷基;C1~C6烷氧基取代的C1~C18烷基。
上述式A化合物、其药学上可接受的盐或前药中:
可选地,式I中,
虚线表示没有化学键或为单键;
G1~G8如权利要求4中所述式I化合物中的定义;
R3选自:C1~C6烷基,所述C1~C6烷基被吗啉基、哌啶基、哌嗪基、吡啶基、苯基或-NH-CO-C1~C6亚烷基(NH2)(A1)中至少一种取代,所述吗啉基、哌啶基、哌嗪基、吡啶基、苯基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;C2~C6烷基,所述C2~C6烷基被羟基取代;C3~C6烷基,所述C3~C6烷基被-NR13R14取代;
其中A1选自:-H;C1~C6烷基,所述C1~C6烷基任选被咪唑基、吲哚基取代,所述咪唑基、吲哚基任选被氨基、羟基、卤素、C1~C6烷基、C1~C6卤代烷基取代;R13、R14各自独立地选自:-H;C1~C6烷基;
R1和R2各自独立地选自C1~C6烷基,所述C1~C6烷基被苯基、萘基、吡啶基中的至少一种取代;
可选地,
式I中,
虚线表示没有化学键或为单键;
G1~G8各自独立地选自-H、卤素、烯丙基、异戊烯基;
R3选自-H;-Me;-(CH2)1~6OH;-(CH2)0~6NH2;-(CH2)1~4NMe2;-(CH2)1~4CN;-(CH2)1~ 4CO2H;-(CH2)1~4C6H5OH;-(CH2)1~4C6H5OMe;-(CH2)1~4C6H5NH2;-(CH2)1~4M;吗啉乙基;哌啶乙基;哌嗪乙基;甲基哌嗪乙基;吡啶乙基;卤代苯乙基;-(CH2)1~6-NH-CO-CH(NH2)(A1);其中,A1、R13、R14如权利要求4中所述,M为单糖基;
或者,所述R3选自吗啉乙基;哌啶乙基;哌嗪乙基;甲基哌嗪乙基;吡啶乙基;氨基苯甲基;氨基苯乙基;羟基苯甲基;卤代苯乙基;-(CH2)1~6-NH-CO-CH(NH2)(A1);-(CH2)2~ 4OH;-(CH2)3~6NH2;-(CH2)1~4NMe2;其中,A1选自-H;咪唑甲基;吲哚甲基;
或者,所述R3选自2-(4-吗啉基)乙基;2-(哌啶-1-基)乙基;2-(哌嗪-1-基)乙基;2-(4-甲基哌嗪-1-基)乙基;2-(吡啶-2-基)乙基;对氨基苯甲基;对氨基苯乙基;对羟基苯甲基;2-(2-氯-6-氟苯基)乙基;-(CH2)1~6-NH-CO-CH(NH2)(A1);-(CH2)2~4OH;-(CH2)3~6NH2;-(CH2)1~4NMe2;其中,A1选自-H;(咪唑-4-基)甲基;(吲哚-3-基)甲基;
R1和R2各自独立地选自-H;-Et;-(CH2)1~4CN;-(CH2)1~4CO2H;-(CH2)1~4OH;苯乙基;萘乙基;吡啶乙基;
或者,所述R1和R2各自独立地选自苯乙基;萘乙基;吡啶乙基;
可选地,式II中,
G1~G8各自独立地选自-H;卤素;
R3为-H;
R4与R5一起构成=O,和/或R6与R7一起构成=O;
当R4与R5或R6与R7不构成=O时,各自独立地选自-H或-OR35,R35为-H或者C1~C6烷基;
R9、R10均为甲基;
R8选自-C(=Y1)-Y2;-S(=O)2-Y3;
其中,Y1选自=O;=S;=NH;
Y2选自:羟胺基;苯基,所述苯基任选被卤素、C1~C6卤代烷基取代;咪唑基;氧代吲哚基乙氨基;氧代苯基乙氨基;(吗啉乙基)哌嗪基;(卤代苯基)甲氨基;(卤代苯基)乙氨基;(卤代甲基苯基)乙氨基;苯基甲氨基;(甲氧基苯基)甲氨基;羟丙氨基;4-(N,N-双(2-氯乙基)氨基)苯基丙基;
或者,Y2选自:羟胺基;苯基;卤代苯基;三氟甲基取代的苯基;2-氧亚基-2-(1H-吲哚-3-基)-1-乙氨基;咪唑-1-基;2-氧亚基-2-苯基-1-乙氨基;4-(2(吗啉-1-基)乙基)哌嗪-1-基;(2,6-二氟苯基)甲氨基;(3-氯-4-氟苯基)甲氨基;2-(2-氯-6-氟苯基)-1-乙氨基;2-(4-三氟甲基苯基)-1-乙氨基;苯基甲基氨基;(4-甲氧基苯基)甲氨基;(S)-2-羟基-1-丙氨基;4-(N,N-双(2-氯乙基)氨基)苯基丙基;
Y3为任选被卤素、C1~C6卤代烷基取代的苯基;
或者,Y3选自卤代苯基;三氟甲基取代的苯基;
或者,
R9与R10一起构成-C(=O)-;
R8选自-H;C1~C18烷基;C1~C6烷氧基取代的C1~C18烷基;或者,R8为甲基。
可选地,上述式A化合物、其药学上可接受的盐或前药,其选自下列化合物、其药学上可接受的盐或前药:
可选地,上述式A化合物、其药学上可接受的盐或前药中,所述药学上可接受的盐包括有机或无机酸的盐;可选地,所述药学上可接受的盐选自所述式A化合物与以下酸化合物形成的盐:盐酸;硫酸;磷酸;甲酸;乙酸;丙酸;乳酸;柠檬酸;酒石酸;琥珀酸;富马酸;马来酸;杏仁酸;苹果酸;樟脑磺酸;
可选地,所述药学上可接受的前药包括所述式A化合物的磷酸酯前药或氨基甲酸酯前药。
本发明还提供一种制备上述式A化合物、其药学上可接受的盐或前药的方法,其特征在于:包括式I-1化合物、I-2化合物、I-3化合物、II-1化合物、或者II-2化合物的制备步骤,
所述式I-1化合物中,G1~G8、R1、R2如上所述,所述式I-1化合物的制备步骤包括:
1)式a3化合物的制备步骤,选自以下方法(1)或方法(2),
方法(1):将式a1化合物和式a2化合物通过Perkin缩合反应制得式a3化合物;
方法(2):将式a2化合物、式a6化合物以及式a7化合物通过Grignard反应,并与碘乙烷反应制得式a8化合物,再将式a8化合物通过在碱性条件下水解,然后酸化制得式a3化合物;
和2)式I-1化合物的制备步骤,
由式a3化合物通过反应制得式I-1化合物;
所述式I-2化合物中,G1~G8、R1、R2、R3如上所述,但是R3不为H,所述式I-2化合物的制备步骤包括:由式I-1化合物制得式I-2化合物;
所述式I-3化合物中G1~G8、R1、R2、R3如上所述,所述式I-3化合物的制备步骤包括:
1):将式I-1化合物通过环化反应或者二氯二氰对苯醌(DDQ)氧化环化反应制得式a5化合物,再由式a5化合物制得式I-3化合物,其中,式I-3化合物中R3为H;
或者2):将式I-2化合物通过光照环化反应或者二氯二氰对苯醌(DDQ)氧化环化反应制得式I-3化合物,其中,式I-3化合物中R3不为H;
其中,当R1、R2为活泼基团时任选采用保护基团进行保护;
所述式II-1化合物中,R9、R10为-H或烷基;G1~G8、R3、R4、R5、R6、R7、R8如上所述,所述式II-1化合物的制备步骤包括:II-1-A类化合物、II-1-B类化合物、II-1-C类化合物、II-1-D类化合物或者II-1-E类化合物的制备步骤,
所述II-1-A类化合物为如下定义的式II-1化合物:式II-1化合物中R8为-C(=Y1)-Y2,Y1选自=O或=S;Y2为芳基,所述芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;
所述II-1-A类化合物制备步骤包括:由式b1化合物与芳基甲酰试剂或芳基磺酰试剂通过酰化反应制得,所述芳基甲酰试剂或芳基磺酰试剂中的芳基上的取代基与Y2芳基上的取代基相同;所述式b1化合物中,G1~G8、R3、R4、R5、R6、R7、R9、R10如II-1-A类化合物中所述;
所述II-1-B类化合物为如下定义的式II-1化合物:式II-1化合物中R8为-C(=Y1)-Y2,Y1选自=O或=S;Y2为咪唑基;所述II-1-B类化合物制备步骤包括:由式b1化合物与1,1′-硫代羰基二咪唑或者三光气与咪唑反应得到式b2化合物;其中所述式b2化合物中Y1为=O或=S;
所述II-1-C类化合物为如下定义的式II-1化合物:式II-1化合物中R8为-C(=Y1)-Y2,Y1选自=O或=S;Y2选自羟胺基;-NR13R14;脂杂环基取代的烷基取代的脂杂环基,所述的脂杂环基均任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;烷氨基,所述烷氨基被氧、羟基、杂芳基、芳基中至少一种取代,所述杂芳基、芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;其中,R13、R14如上所述;所述II-1-C类化合物的制备步骤包括:式b2化合物与碘甲烷反应生成式b3化合物,式b3化合物与化合物R反应制得;其中,所述化合物R选自脂杂环基取代的烷基取代的脂杂环,所述的脂杂环基和脂杂环均任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;烷氨,所述烷氨被氧、羟基、杂芳基、芳基中至少一种取代,所述杂芳基、芳基任选被氨基、羟基、卤素、烷氧基、烷基、卤代烷基取代;
所述II-1-D类化合物为如下定义的式II-1化合物:式II-1化合物中R8为-C(=NH)-NHOH;所述II-1-D类化合物的制备步骤包括:由式b1化合物制得式b4化合物,式b4化合物与盐酸羟胺反应制得;
所述II-1-E类化合物为如下定义的式II-1化合物:式II-1化合物中R8为-C(=Y1)-Y2,Y1为=O,Y2为4-(N,N-双(2-氯乙基)氨基)苯基丙基;所述II-1-E类化合物的制备步骤包括:式b1化合物与苯丁酸氮芥反应制得;
所述式II-2化合物中,R9与R10一起构成-C(=O)-;G1~G8、R3、R4、R5、R6、R7、R8如上所述;所述式II-2化合物的制备步骤包括:式c1化合物与式c2化合物反应制得式c3化合物,再通过反应制得式c4化合物,再通过反应制得式c5化合物,再通过反应制得式c6化合物,再通过反应制得式c7化合物,再通过反应制得式c8化合物,再通过反应制得式c9化合物,再通过反应制得式II-2化合物;
R3为活泼基团时任选地采用保护基团进行保护;
可选地,所述Perkin缩合反应是在草酰氯、三乙胺(Et3N)、二氯甲烷的存在下进行的;
可选地,所述氨解反应是在六甲基二硅胺脘(HMDS)、N,N-二甲基甲酰胺和甲醇的存在下进行的;
可选地,所述光照环化反应中采用丙酮作为溶剂,采用碘单质作为催化剂,在高压汞灯光照下进行;
可选地,所述二氯二氰对苯醌(DDQ)氧化环化反应是在对甲基苯磺酸的催化下,在苯溶剂中与DDQ发生氧化关环反应;
可选地,所述式a6化合物通过二溴马来酰亚胺与碘甲烷反应制得;
可选地,所述式a8化合物通过在KOH或NaOH等碱性溶液中水解,经盐酸等酸化后得到式a3化合物;
可选地,当R1、R2或R3为活泼基团时,所述保护基团选自(Boc)2O;
可选地,所述II-1-A类化合物中R3为-H;R6与R7一起构成=O;R8为甲基,所述II-1-A类化合物的制备步骤包括:①酰化反应,将十字孢碱溶于二氯甲烷,加入三乙胺,与卤代苯甲酰化试剂发生酰化反应;②卤代反应,将步骤①所得产物在甲醇中与卤代丁二酰亚胺室温下发生卤代反应;③氧化反应,将步骤②所得产物用氧化剂氧化得到含R4、R5独立选自-H;-OH;或R4与R5一起构成=O的化合物;所述酰化反应、卤代反应、氧化反应按任选的顺序进行;可选地,所述氧化反应采用下述试剂进行:O2、DMSO、t-BuOK和任选的NaOH;
可选地,所述II-1-B类化合物中,式b2化合物中的R3为-H;R4、R5独立选自-H;R6与R7一起构成=O;R8为甲基;所述II-1-B类化合物的制备通过以下方法进行:方法①:将十字孢碱溶于二氯甲烷,然后加入三乙胺,与1,1′-硫代羰基二咪唑反应制得;或者方法②:将十字孢碱溶于四氢呋喃,然后加入二异丙基乙胺和三光气反应,将反应粗产物溶于四氢呋喃,加入二异丙基乙胺、咪唑和对二甲氨基吡啶制得;
可选地,所述II-1-C类化合物中R3为-H;R4、R5均为-H;R6与R7一起构成=O;R8为甲基;所述II-1-C类化合物的制备步骤包括:将式b2化合物在乙腈溶剂中与碘甲烷成盐,后溶于二氯甲烷,加入三乙胺和化合物R进行反应取代咪唑盐制得;
可选地,所述II-1-D类化合物中R3为-H;R4、R5均为-H;R6与R7一起构成=O;R8为甲基;所述分类(4)化合物的制备方法包括以下步骤:Fradcarbazole C与盐酸羟胺反应制得;
可选地,所述II-1-E类化合物中R3为-H,R4、R5均为-H;R6与R7一起构成=O;R8为甲基;所述II-1-E类化合物的制备步骤包括:十字孢碱与苯丁酸氮芥反应制得;
可选地,所述式II-2化合物中,R3为-H;R4与R5一起构成=O;R6与R7一起构成=O;R8为甲基;所述式II-2化合物的制备步骤包括:①以葡萄糖为原料,经过全乙酰化、1-位溴代、1,2-位成烯反应、脱乙酰基、6-位羟基的TIPS保护、3,4-位构建噁唑环酮,甲基化,羟汞化硼氢化钠还原在1-位引入羟基即得到糖供体;②以2,3-二溴马来酰亚胺为原料,经过BOM保护,与吲哚格氏试剂反应引入一分子吲哚,再将吲哚的氮氢用Boc保护,之后再与吲哚格氏试剂反应得到母核;③将步骤①所述糖供体与步骤②所述母核,利用Mitsunobu反应进行糖苷化形成第一个糖苷键,糖苷键的异构体通过硅胶柱色谱进行分离,然后进行Boc和TIPS保护基的脱除,高压汞灯照射进行合环,再将6-位羟基以碘取代,5,6-位脱碘成双键,再在碘催化下形成第二个糖苷键,用四丁基氢化锡脱碘,20%的氢氧化钯碳脱除BOM制得;
可选地,所述式II-2化合物中,R3为-H;R8为甲基;R4与R5一起构成=O,R6、R7均为-H;或者,R4、R5均为-H,R6与R7一起构成=O;所述式II-2化合物的制备步骤包括:将所述R3为-H,R4与R5一起构成=O,R6与R7一起构成=O,R8为甲基的式II-2化合物经过硼氢化钠还原和锌粉醋酸还原制得;
可选地,所述式II-2化合物采用以D-葡萄糖或L-葡萄糖进行糖苷键及噁唑环酮构型不相同的异构体的制备。
本发明中,上述化合物1~81、158~166及其重要中间产物24c、26c、43c、46c、49c、52c和58c可以从吲哚或卤素取代的吲哚和吲哚乙酸经下列化学合成方法制备而得:
上述式A-I、式A-II中,G1~G8各自独立地为-H、-F、-Cl、-Br、烯丙基或异戊烯基;X为-H、-Me、-NH2、2-(4-吗啉基)乙基、2-(哌啶-1-基)乙基、-(CH2)nNH2、-(CH2)nNMe2、-(CH2)nCN、-(CH2)nCO2H、-(CH2)nOY、-(CH2)nC6H5R3(Y为-H、葡萄糖基或氨基酸基,n=1~4,R3=-OH、-OMe);R1,R2代表-H、-Et、-(CH2)mCN、-(CH2)mCO2H、-(CH2)mOH(m=1~4)、-CH2Ph。
化合物82~101可以由卤素取代的吲哚和二溴马来酰亚胺经下列化学合成方法制备而得:
上述式A-III中,G5~G8各自独立地为-H、-F、-Cl或-Br;X为-H、-Me或-CH2OH,R1,R2各自独立地为-H、-Et或-CH2OH。
化合物102~129及其中间产物102f和116c可以从吲哚和二溴马来酰亚胺经下列化学合成方法制备而得:
上述式A-IV中,G5~G8均为-H;X代表-H、-Me、2-(哌嗪-1-基)乙基、2-(2-氯-6-氟-苯-1-基)乙基、2-(4-吗啉基)乙基、2-(哌啶-1-基)乙基、2-(4-甲基哌嗪-1-基)乙基、-(CH2)nOH、-(CH2)nNH2、(n=1,2);R1为苯基乙基或2-(萘-1-基)乙基。
化合物130~157可由十字孢碱(staurosporine)与含卤素苯甲酰试剂和苯磺酰试剂经下列化学合成方法制备而得:
上述式A-V和A-VI中,R1为-H、-F、-Cl、-Br、-I或-CF3;R2、R3各自独立地为=O、-H或-OH;X为-H或-Br。
化合物167~184可由十字孢碱与硫羰基二咪唑或三光气、咪唑、碘甲烷以及胺类化合物制备;或由十字孢碱与苯丁酸氮芥经下列化学合成方法制备:
上述式A-VII中,R4代表=O、=S或=NH,R5代表2-氧亚基-2-(1H-吲哚-3-基)-1-乙氨基、咪唑-1-基、羟胺基、2-氧亚基-2-苯基-1-乙氨基、4-(2-(4-吗啉基)乙基)哌嗪-1-基、(2,6-二氟苯基)甲氨基、(3-氯-4-氟苯基)甲氨基、2-(2-氯-6-氟苯基)-1-乙氨基、2-(3-三氟甲基苯基)-1-乙氨基、苯基甲基氨基、(4-甲氧基苯基)甲氨基、(S)-2-羟基-1-丙氨基、4-(N,N-双(2-氯乙基)氨基苯基)丙基。
式A-VIII中的化合物为185~196,其中化合物185~187可由D-葡萄糖和二溴马来酰亚胺经下列化学合成方法制备而得,式A-VIII中,188~190可由化合物26b以相同的方法制得;191~195可由L-葡萄糖替代D-葡萄糖以相同的方法制得。
上述式A-VIII中,R1a、R1b各自独立地为-H或-OH,或者R1a、R1b一起形成=O,R2a、R2b各自独立地为-H或-OH,或者R2a、R2b一起形成=O,R3为-H、-Me、2-(哌嗪-1-基)乙基、2-(2-氯-6-氟苯基)乙基、2-(4-吗啉基)乙基、2-(哌啶-1-基)乙基、2-(4-甲基哌嗪-1-基)乙基、-(CH2)nOH或-(CH2)nNH2,(n=1或2);R3代表H、-(CH2)nCH3(n=0~17)或-(CH2)nOY,(n=1,2、Y=Me,Et)。
本发明还提供一种α-葡萄糖苷酶抑制组合物,包括上述式A化合物、其药学上可接受的盐或前药中的至少一种和药学上可接受的辅料。
本发明还提供一种抗糖尿病的药物组合物,包括上述式A化合物、其药学上可接受的盐或前药中的至少一种和药学上可接受的辅料。可选地,所述抗糖尿病药物为抗α-葡萄糖苷酶介导的糖尿病的药物。
本发明还提供一种抗糖尿病并发症的药物组合物,包括上述式A化合物、其药学上可接受的盐或前药中的至少一种和药学上可接受的辅料。可选地,所述糖尿病并发症为糖尿病肾病。
本发明式A化合物、其药学上可接受的盐或前药、α-葡萄糖苷酶抑制组合物、或者抗糖尿病药物组合物可以通过口服给药和注射给药,也适合其它的给药方式,如经皮给药等。本发明的抗糖尿病药物组合物可以为片剂、胶囊、粉剂、颗粒、锭剂、栓剂、口服液或无菌胃肠外悬液等制剂形式。还包括各种容量的注射剂、冻干粉剂等针剂形式。上述剂型的药物均可按照药学领域的常规方法制备。本发明抗糖尿病药物组合物中,所用的辅料包括本领域常规的辅料,例如稀释剂、填充剂、粘合剂、湿润剂、吸收促进剂、表面活性剂、吸附载体、润滑剂等。
本发明还提供上述式A化合物、其药学上可接受的盐或前药在制备α-葡萄糖苷酶抑制剂中的用途。
本发明还提供上述式A化合物、其药学上可接受的盐或前药在制备抗糖尿病药物中的用途,可选地,所述抗糖尿病药物为抗α-葡萄糖苷酶介导的糖尿病的药物。
本发明还提供上述式A化合物、其药学上可接受的盐或前药在制备抗糖尿病并发症药物中的用途,可选地,所述糖尿病为α-葡萄糖苷酶介导的糖尿病。可选地,所述糖尿病并发症为糖尿病肾病。
本发明还提供上述式A化合物、其药学上可接受的盐或前药用作为α-葡萄糖苷酶抑制剂的用途。
本发明还提供上述式A化合物、其药学上可接受的盐或前药用于抗糖尿病的用途。可选地,所述糖尿病为α-葡萄糖苷酶介导的糖尿病。
本发明还提供上述式A化合物、其药学上可接受的盐或前药用于抗糖尿病并发症的用途,可选地,所述并发症为糖尿病肾病。
有益效果
本发明的化合物具有较强的α-葡萄糖苷酶抑制作用,具有良好的抗糖尿病以及抗糖尿病并发症如糖尿病肾病的活性。
附图说明
图1:显示化合物49对糖尿病肾病鼠的治疗作用的肾皮质组织切片H&E染色显微照片:×400。
A为正常组,B为db/db模型组,C为模型组+氯沙坦组(阳性药),D为模型组+化合物49(0.5mg/kg/day)组,E为模型组+化合物49(1.0mg/kg/day)组,F为模型组+化合物49(2.0mg/kg/day)组。
具体实施方式:
以下通过实施例对本发明进行示例性说明,但本发明的范围不限于这些实施例。
【实施例1】化合物1~196的制备
化合物1的制备
i)1-乙基-3-吲哚乙酸(1a)的制备
氩气保护下,在250mL三口瓶中加入4g氢化钠(质量分数60%,分散于石蜡中)、80mL四氢呋喃,于0℃搅拌悬浮,加入30mL四氢呋喃溶解的吲哚-3-乙酸(3.5g,20mmol),搅拌半小时后,滴加30mL四氢呋喃溶解的碘乙烷(5mL,60mmol),缓慢升至室温,反应过夜后,降至0℃下,滴加10滴甲醇,再加适量水至得亮黄色溶液,乙酸乙酯萃取,水层加浓盐酸后再萃取,合并有机层,并用无水Na2SO4干燥,真空蒸干,后经过硅胶柱色谱分离、石油醚:乙酸乙酯=8:1(v/v)洗脱得产物(1a)3.87g,收率95.4%。1H NMR(600MHz,CDCl3)δ7.59(d,1H,J=8.2Hz,Ar-H),7.30(d,1H,J=8.2Hz,Ar-H),7.20(dt,1H,J=8.2Hz,0.9Hz,Ar-H),7.10(dt,1H,J=8.2Hz,1.0Hz,Ar-H),7.07(s,1H,Ar-H),4.10(q,2H,J=7.3Hz,CH3-CH 2-),3.77(s,2H,-CH 2-CO2H),1.41(t,3H,J=7.3Hz,-CH2-CH 3).13C NMR(150MHz,CDCl3)δ178.6,135.9,127.6,126.1,121.7,119.2,119.0,109.4,106.0,40.8,31.1,15.4.ESI-MS m/z 202.1[M-H]–.
ii)1-氰甲基吲哚(1b)的制备
氩气保护下,在100mL三口瓶中加入180mg氢化钠(质量分数60%,分散在石蜡中)和30mL乙腈,于-5℃搅拌悬浮,滴加10mL乙腈溶解的吲哚(351mg,3.0mmol),搅拌反应30min,缓慢滴加10mL乙腈溶解的2-溴代乙腈(300μL,4.5mmol),缓慢升至室温,反应24h后,加入饱和NH4Cl水溶液终止反应,乙酸乙酯萃取(100mL×3次),合并有机层,并用无水Na2SO4干燥,真空蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=15:1(v/v)洗脱得无色油状产物(1b)299mg,收率64%。1H NMR(600MHz,CDCl3)δ7.64(d,1H,J=8.2Hz,Ar-H),7.33-7.28(m,2H,Ar-H),7.19(d,1H,J=6.9Hz,Ar-H),7.02(d,1H,J=3.2Hz,Ar-H),6.58(d,1H,J=3.2Hz,Ar-H),4.87(s,2H,-CH 2-CN).13C NMR(150MHz,CDCl3)δ135.6,128.9,127.1,122.8,121.5,120.8,114.4,108.7,104.1,34.1.ESI-MS m/z 157.1[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酸酐(1c)的制备
将化合物1a(299mg,1.92mmol)用25mL CH2Cl2溶解,在-5℃下滴加5mL CH2Cl2溶解的草酰氯(366mg,2.88mmol),滴毕,在-5℃反应2h,后补加草酰氯0.5mL,继续反应2.5h,升至室温,真空抽干溶剂得黄色晶体。用30mL CH2Cl2重新溶解,滴加到15mL CH2Cl2溶解的化合物1b(390mg,1.92mmol)和三乙胺(388mg,3.84mmol)中,经硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末(1c)270mg,收率36%。1H NMR(600MHz,DMSO-d6)δ7.97(s,1H,Ar-H),7.95(s,1H,Ar-H),7.64(d,1H,J=8.2Hz,Ar-H),7.54(d,1H,J=8.2Hz,Ar-H),7.22(t,1H,J=7.3Hz,Ar-H),7.12(t,1H,J=7.8Hz,Ar-H),6.90(d,1H,J=7.7Hz,Ar-H),6.87(d,1H,J=7.8Hz,Ar-H),6.85(t,1H,J=7.7Hz,Ar-H),6.77(t,1H,J=7.8Hz,Ar-H),5.67(s,2H,-CH 2-CN),4.28(q,2H,J=7.3Hz,-CH 2-CH3),1.33(t,3H,J=7.3Hz,-CH2-CH 3).13C NMR(150MHz,DMSO-d6)δ166.2,166.1,135.7,135.6,133.2,132.5,129.9,126.1,125.5,125.3,123.1,122.3,121.7,121.6,120.9,120.3,116.0,110.6,110.4,106.0,104.0,40.9,34.1,15.2.ESI-MS m/z 396.2[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(1)的制备
在单口瓶中,用4mL DMF溶解化合物1c(126mg,0.32mmol),密封后搅拌下将HMDS(6.7mL,32mmol)和MeOH(0.64mL,16mmol)混合后,注入到单口瓶中,反应液即由红色变为淡黄色混浊物,随着反应进行,渐变为澄清液,颜色渐变为橙红色。反应过夜后,倒入25mL冷水中,乙酸乙酯萃取(50mL×3次),合并有机层,并用无水Na2SO4干燥,真空蒸干。硅胶柱色谱分离、氯仿洗脱得橙红色粉末(1)118mg,收率94%。1H NMR(600MHz,DMSO-d6)δ11.00(s,1H,imide-NH),7.87(s,1H,Ar-H),7.82(s,1H,Ar-H),7.57(d,1H,J=8.2Hz,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.15(t,1H,J=8.3Hz,Ar-H),7.05(t,1H,J=7.4Hz,Ar-H),6.83(d,1H,J=8.3Hz,Ar-H),6.80(d,1H,J=8.2Hz,Ar-H),6.75(t,1H,J=7.8Hz,Ar-H),6.68(t,1H,J=7.8Hz,Ar-H),5.64(s,2H,N-CH 2-CN),4.26(q,2H,J=7.3Hz,-CH 2-CH3),1.32(t,3H,J=7.3Hz,-CH2-CH 3).13C NMR(150MHz,DMSO-d6)δ172.7,172.6,135.5,135.4,131.8,131.4,129.1,126.0,125.8,122.6,121.8,121.5,121.2,121.0,120.4,119.6,116.1,110.1,110.0,106.8,104.6,40.7,34.0,15.2.ESI-MS m/z 395.2[M+H]+.
化合物2的制备
i)1-氰乙基吲哚(2b)的制备
按照化合物1b的制备方法,氢化钠(360mg,9.0mmol,分散在石蜡中,质量分数60%)、吲哚(702mg,6.0mmol)和3-溴代丙腈(744μL,9.0mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=15:1(v/v)洗脱得无色油状产物(2b)949mg,收率94%。1H NMR(600MHz,CDCl3)δ7.65(d,1H,J=7.8Hz,Ar-H),7.30(d,1H,J=7.8Hz,Ar-H),7.25(t,1H,J=7.8Hz,Ar-H),7.15(t,1H,J=6.8Hz,Ar-H),7.13(d,1H,J=3.2Hz,Ar-H),6.55(d,1H,J=3.2,Ar-H),4.42(t,2H,J=6.9Hz,N-CH 2-CH2CN),2.78(t,2H,J=J=6.9Hz,NCH2-CH2 -CN).13C NMR(150MHz,CDCl3)δ135.4,129.1,127.5,122.3,121.6,120.2,117.4,108.7,103.0,42.2,19.2.
ii)2-(1-乙基-3-吲哚)-3-(1-氰乙基-3-吲哚)马来酸酐(2c)的制备
按照化合物1c的制备方法,由化合物2b(988mg,5.81mmol)、草酰氯(1107mg,8.72mmol)、1a(1180mg,5.81mmol)和三乙胺(1176mg,11.64mmol)制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得红色粉末(2c)530mg,收率23%。1H NMR(600MHz,CDCl3)δ7.84(s,1H,Ar-H),7.66(s,1H,Ar-H),7.35(d,1H,J=8.3Hz,Ar-H),7.34(d,1H,J=8.3Hz,Ar-H),7.22(dt,1H,J=7.3Hz,1.0Hz,Ar-H),7.16(dt,1H,J=7.5Hz,1.4Hz,Ar-H),7.11(d,1H,J=7.8Hz,Ar-H),6.90(dt,1H,J=7.8Hz,1.0Hz,Ar-H),6.89(d,1H,J=8.2Hz,Ar-H),6.82(dt,1H,J=7.3Hz,1.0Hz,Ar-H),4.47(t,2H,J=6.9Hz,N-CH 2-CH2-CN),4.23(q,2H,J=7.3Hz,-CH 2-CH3),2.85(t,2H,J=6.9Hz,-NCH2-CH 2-CN),1.51(t,3H,J=7.3Hz,-CH2-CH 3).13C NMR(150MHz,CDCl3)δ166.7,166.6,136.2,135.5,132.8,131.5,129.6,126.2,126.0,125.6,123.4,122.9,122.8,122.5,121.3,120.8,116.5,110.0,109.1,106.7,105.0,42.4,41.7,19.0,15.1.ESI-MS m/z 410.2[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-氰乙基-3-吲哚)马来酰亚胺(2d)的制备
按照化合物1的制备方法,由化合物2c(230mg,0.56mmol)、HMDS(12mL,57mmol)和MeOH(1.2mL,28.5mmol)制备,经硅胶柱色谱分离、氯仿洗脱得橙红色粉末(2d)219mg,收率95%。1H NMR(600MHz,CDCl3)δ7.75(s,1H,Ar-H),7.58(s,1H,Ar-H),7.51(s,1H,-NH),7.31(d,1H,J=8.2Hz,Ar-H),7.29(d,1H,J=8.2Hz,Ar-H),7.16(t,1H,J=7.3Hz,Ar-H),7.11(t,1H,J=8.2Hz,Ar-H),7.09(d,1H,J=8.2Hz,Ar-H),6.88(d,1H,J=8.2Hz,Ar-H),6.85(t,1H,J=7.4Hz,Ar-H),6.77(t,1H,J=7.3Hz,Ar-H),4.45(t,2H,J=6.9Hz,N-CH 2-CH2CN),4.20(q,2H,J=7.3Hz,-CH 2-CH3),2.80(t,2H,J=6.9Hz,-NCH2-CH 2-CN),1.48(t,3H,J=7.3Hz,-CH2CH 3).13C NMR(150MHz,CDCl3)δ171.8,171.7,136.0,135.4,131.7,130.6,129.5,126.6,126.3,125.9,123.0,122.7,122.3,122.2,120.8,120.2,116.7,109.6,108.8,107.3,105.4,42.3,41.5,18.9,15.2.ESIMS:m/z 407.1[M–H]–.
iv)N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-氰乙基-3-吲哚)马来酰亚胺(2)的制备
在15mL反应瓶中加入化合物2d(16.2mg,39.7μmol)和NaHCO3(6.7mg,79.4μmol),加入甲醛溶液(3mL,质量分数37%),85℃搅拌反应10小时后倒入冷水中。乙酸乙酯萃取,合并有机层,并用无水Na2SO4干燥,真空蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末(2)17.1mg,收率98%。1H NMR(600MHz,DMSO-d6)δ7.93(s,1H,Ar-H),7.85(s,1H,Ar-H),7.59(d,1H,J=8.2Hz,Ar-H),7.47(d,1H,J=8.3Hz,Ar-H),7.06(dt,1H,J=7.8Hz,0.9Hz,Ar-H),7.04(dt,1H,J=7.3Hz,1.0Hz,Ar-H),6.83(d,1H,J=8.3Hz,Ar-H),6.78(d,1H,J=8.3Hz,Ar-H),6.69(dt,1H,J=7.3Hz,1.0Hz,Ar-H),6.68(dt,1H,J=7.8Hz,0.9Hz,Ar-H),6.31(t,1H,J=7.0Hz,-OH),4.98(d,2H,J=6.9Hz,N-CH 2-OH),4.59(t,2H,J=6.4Hz,N-CH 2-CH2CN),4.26(q,2H,J=7.3Hz,CH 2-CH3),3.03(t,2H,J=6.4Hz,N-CH2-CH 2-CN),1.34(t,3H,J=7.3Hz,-CH2CH 3).13C NMR(150MHz,DMSO-d6)δ171.5,171.4,136.2,136.1,132.3,132.2,128.4,126.7,126.5,126.4,122.6,122.3,121.9,121.7,120.5,120.3,119.0,110.9,110.7,106.2,105.3,60.8,42.0,41.3,19.1,15.8.ESI-MS m/z 439.2[M+H]+.
化合物3的制备
i)1-氰丙基吲哚(3b)的制备
按照化合物1b的制备方法,由吲哚(1170mg,10mmol)、NaH(600mg,15mmol,质量分数60%,分散在石蜡中)和4-溴代丁腈(1.6mL,15mmol)制备,硅胶柱色谱分离、石油醚:乙酸乙酯=20:1(v/v)洗脱得无色油状产物(3b)1042mg,收率57%。1H NMR(600MHz,CDCl3)δ7.70(d,1H,J=7.7Hz,Ar-H),7.38(d,1H,J=8.3Hz,Ar-H),7.29(t,1H,J=7.7Hz,Ar-H),7.19(t,1H,J=7.4Hz,Ar-H),7.12(d,1H,J=3.3Hz,Ar-H),6.58(d,1H,J=3.3Hz,Ar-H),4.28(t,2H,J=6.6Hz,N-CH2 -(CH2)2CN),2.19(t,2H,J=5.5Hz,N(CH2)2-CH2 -CN),2.16-2.14(m,2H,-NCH2-CH2 -CH2CN)。13C NMR(150MHz,CDCl3)δ135.9,128.9,127.8,122.1,121.4,119.9,119.0,109.3,102.2,44.5,26.1,14.7.ESI-MS m/z 185.1[M+H]+.
ii)2-(1-乙基-3-吲哚)-3-(1-氰丙基-3-吲哚)马来酸酐(3c)的制备
按照化合物1c的制备方法,由3b(407mg,2.21mmol)、草酰氯(421mg,3.32mmol)、化合物1a(449mg,2.21mmol)和三乙胺(447mg,4.42mmol)制备,硅胶柱色谱分离、氯仿洗脱得红色粉末(3c)415mg,收率45%。1H NMR(600MHz,DMSO-d6)δ7.89(s,1H,Ar-H),7.88(s,1H,Ar-H),7.57(d,1H,J=8.8Hz,Ar-H),7.54(d,1H,J=8.2Hz,Ar-H),7.14(t,1H,J=8.2Hz,Ar-H),7.12(t,1H,J=7.7Hz,Ar-H),6.94(d,1H,J=8.3Hz,Ar-H),6.91(d,1H,J=8.3Hz,Ar-H),6.80(t,1H,J=7.1Hz,Ar-H),6.79(t,1H,J=7.1Hz,Ar-H),4.33(t,2H,J=7.1Hz,N-CH 2-(CH2)2CN),4.26(q,2H,J=7.1Hz,-CH 2-CH3),2.44(t,2H,J=7.4Hz,N(CH2)2-CH 2-CN),2.06-2.04(m,2H,-NCH2-CH 2-CH2CN),1.32(t,3H,J=7.1Hz,-CH2-CH 3)。13C NMR(150MHz,DMSO-d6)δ166.9×2,136.6,136.3,133.6,133.5,128.8,127.8,125.8,125.7,122.9,122.8,122.3,122.2,120.8,120.7,120.4,111.1,111.0,105.2,104.7,55.5,40.4,26.1,15.7,14.3.ESI-MS m/z424.2[M+H]+.
ⅲ)2-(1-乙基-3-吲哚)-3-(1-氰丙基-3-吲哚)马来酰亚胺(3)的制备
按照化合物1的制备方法,由3c(205mg,0.49mmol)、HMDS(10.2mL,48.5mmol)和MeOH(0.97mL,24.3mmol)制备,硅胶柱色谱分离、氯仿洗脱得橙红色粉末(3)203mg,收率98%。1H NMR(600MHz,DMSO-d6)δ10.94(s,1H,imide-NH),7.78(s,1H,Ar-H),7.74(s,1H,Ar-H),7.50(d,1H,J=8.3Hz,Ar-H),7.47(d,1H,J=8.3Hz,Ar-H),7.07(dt,1H,J=7.1Hz,1.1Hz,Ar-H),7.05(dt,1H,J=7.1Hz,1.1Hz,Ar-H),6.86(d,1H,J=7.7Hz,Ar-H),6.85(d,1H,J=7.7Hz,Ar-H),6.71(t,1H,J=7.7Hz,Ar-H),6.69(t,1H,J=7.1Hz,Ar-H),4.29(t,2H,J=6.6Hz,N-CH 2-(CH2)2CN),4.24(q,2H,J=7.2Hz,-CH 2-CH3),2.40(t,2H,J=7.1Hz,N(CH2)2-CH 2-CN),2.05-2.03(m,2H,NCH2-CH 2-CH2CN),1.32(t,3H,J=7.2Hz,-CH2-CH 3).13CNMR(150MHz,DMSO-d6)δ173.4×2,136.4,136.1,132.4,132.2,128.5,127.6,126.3,126.2,122.4,122.3,122.0,121.9,120.5,120.3,120.0,110.7,110.6,105.9,105.3,45.0,41.2,26.2,15.8,14.3.HR-ESIMS m/z 421.1645[M–H]–(calcd.for C26H21N4O2,421.1665).
化合物4的制备
按照化合物2的制备方法,由化合物3(6.3mg,13.9μmol)、NaHCO3(2.3mg,27.9μmol)和甲醛溶液(3mL,质量分数37%)制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-氰丙基-3-吲哚)马来酰亚胺(4)6.8mg,收率99%。1H NMR(600MHz,CDCl3)δ7.74(s,1H,Ar-H),7.50(s,1H,Ar-H),7.33(d,1H,J=5.5Hz,Ar-H),7.32(d,1H,J=5.5Hz,Ar-H),7.17-7.15(m,2H,Ar-H),7.11(t,1H,J=7.3Hz,Ar-H),6.88(d,2H,J=7.3Hz,Ar-H),6.74(t,1H,J=7.3Hz,Ar-H),5.25(d,2H,J=7.8Hz,N-CH 2-OH),4.27(t,2H,J=6.4Hz,N-CH 2-(CH2)2CN),4.20(q,2H,J=7.3Hz,N-CH 2-CH3),3.15(brs,1H,-OH),2.17(t,2H,J=5.9Hz,N(CH2)2-CH 2-CN),2.13-2.11(m,2H,NCH2-CH 2-CH2CN),1.48(t,3H,J=7.3Hz,-CH2-CH 3).13C NMR(150MHz,CDCl3)δ171.8,171.7,136.2,136.1,131.7,131.2,128.5,126.4,126.3,125.9,122.9,122.8,122.4,122.3,120.7,120.2,118.6,109.9,109.3,106.7,105.5,61.9,44.8,41.6,25.9,15.3,14.6.ESI-MS m/z475.1[M+Na]+.
化合物5的制备
i)1-氰丁基吲哚(5b)的制备
按照化合物1b的制备方法,由吲哚(585mg,5mmol)、NaH(300mg,7.5mmol,质量分数60%,分散在石蜡中)和5-溴代戊腈(880μL,7.5mmol)制备,硅胶柱色谱分离、石油醚:乙酸乙酯=10:1(v/v)洗脱得无色油状产物(5b)627mg,收率83%。1H NMR(600MHz,DMSO-d6)δ7.68(d,1H,J=8.0Hz,Ar-H),7.35(d,1H,J=8.4Hz,Ar-H),7.26(t,1H,J=7.7Hz,Ar-H),7.16(t,1H,J=7.3Hz,Ar-H),7.12(d,1H,J=2.7Hz,Ar-H),6.58(d,1H,J=2.7Hz,Ar-H),4.17(t,2H,J=6.8Hz,N-CH 2-(CH2)3CN),2.26(t,2H,J=7.3Hz,N(CH2)3-CH 2-CN),1.98-1.93(m,2H,NCH2-CH 2-(CH2)2CN),1.61-1.54(m,2H,N(CH2)2-CH 2-CH2CN).13C NMR(150MHz,DMSO-d6)δ136.0,128.8,127.7,121.8,121.3,119.6,119.4,109.3,101.7,45.5,29.4,23.0,17.0.
ii)2-(1-乙基-3-吲哚)-3-(1-氰丁基-3-吲哚)马来酸酐(5c)的制备
按照化合物1c的制备方法,由5b(627mg,3.17mmol)、草酰氯(604mg,4.76mmol)、化合物1a(644mg,3.17mmol)和三乙胺(640mg,6.34mmol)制备,硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱,得红色粉末(5c)712mg,收率51%。1H NMR(600MHz,DMSO-d6)δ7.91(s,1H,Ar-H),7.90(s,1H,Ar-H),7.56(d,1H,J=8.3Hz,Ar-H),7.53(d,1H,J=8.3Hz,Ar-H),7.11(dt,1H,J=7.5Hz,1.0Hz,Ar-H),7.10(dt,1H,J=7.8Hz,0.9Hz,Ar-H),6.88(d,1H,J=8.3Hz,Ar-H),6.86(d,1H,J=8.7Hz,Ar-H),6.76(dt,1H,J=7.8Hz,1.0Hz,Ar-H),6.75(dt,1H,J=7.4Hz,0.9Hz,Ar-H),4.30(t,2H,J=6.9Hz,N-CH 2-(CH2)3CN),4.28(q,2H,J=7.3Hz,N-CH 2-CH3),2.51(t,2H,J=7.3Hz,N(CH2)3-CH 2-CN),1.84-1.81(m,2H,NCH2-CH 2-(CH2)2CN),1.50-1.48(m,2H,N(CH2)2-CH 2-CH2CN),1.34(t,3H,J=7.3Hz,-CH2-CH 3).13C NMR(150MHz,DMSO-d6)δ166.9×2,136.5,136.3,133.7,133.3,128.5,127.9,125.9×2,122.8,122.7,122.1×2,121.0,120.7,120.6,111.2,111.1,104.9,104.8,45.7,41.5,29.3,22.7,16.4,15.7.ESI-MS m/z 438.1[M+H]+.
ⅲ)2-(1-乙基-3-吲哚)-3-(1-氰丁基-3-吲哚)马来酰亚胺(5)的制备
以化合物1的制备方法,由5c(616mg,1.14mmol)、HMDS(12mL,57mmol)和MeOH(1.14mL,28.5mmol)制备,硅胶柱色谱分离、氯仿洗脱得橙红色粉末(5)585mg,收率95%。1HNMR(600MHz,DMSO-d6)δ10.92(s,1H,imide-NH),7.80(s,1H,Ar-H),7.77(s,1H,Ar-H),7.50(d,1H,J=8.2Hz,Ar-H),7.47(d,1H,J=8.2Hz,Ar-H),7.04(dt,1H,J=7.4Hz,0.9Hz,Ar-H),7.03(dt,1H,J=7.8Hz,1.0Hz,Ar-H),6.81(d,1H,J=8.2Hz,Ar-H),6.79(d,1H,J=7.8Hz,Ar-H),6.67(dt,1H,J=7.4Hz,0.9Hz,Ar-H),6.65(dt,1H,J=7.8Hz,1.0Hz,Ar-H),4.28(t,2H,J=6.8Hz,N-CH 2-(CH2)3CN),4.26(q,2H,J=7.3Hz,N-CH 2-CH3),2.52(t,2H,J=7.3Hz,N(CH2)3-CH 2-CN),1.82-1.79(m,2H,NCH2-CH 2-(CH2)2CN),1.50-1.47(m,2H,N(CH2)2-CH 2-CH2CN),1.34(t,3H,J=7.3Hz,-CH2CH 3).13C NMR(150MHz,DMSO-d6)δ173.4×2,136.3,136.0,132.4,132.0,128.8,128.2×2,127.7,126.4,122.3,122.2 121.8,121.0×2,120.0,110.8,110.7,105.6,105.4,45.5,41.3,29.3,22.7,16.4,15.8.ESI-MS m/z 437.1[M+H]+.
化合物6的制备
按照化合物2的制备方法,由化合物5(6.7mg,14.4μmol)、NaHCO3(2.4mg,28.8μmol)和甲醛溶液(3mL,质量分数37%)制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=5:2(v/v)洗脱得红色粉末N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-氰丁基-3-吲哚)马来酰亚胺(6)6.9mg,收率96%。1H NMR(600MHz,CDCl3)δ7.70(s,1H,Ar-H),7.55(s,1H,Ar-H),7.32(d,1H,J=8.2Hz,Ar-H),7.27(d,1H,J=8.3Hz,Ar-H),7.13(dt,1H,J=7.8Hz,1.0Hz,Ar-H),7.11(dt,1H,J=7.8Hz,1.0Hz,Ar-H),7.06(d,1H,J=7.8Hz,Ar-H),6.91(d,1H,J=7.8Hz,Ar-H),6.81(d,1H,J=7.8Hz,Ar-H),6.74(d,1H,J=7.8Hz,Ar-H),5.25(d,2H,J=7.8Hz,N-CH 2-OH),4.19(q,2H,J=7.3Hz,N-CH 2-CH3),4.15(t,2H,J=6.4Hz,N-CH 2-(CH2)3CN),3.34(t,1H,J=7.8Hz,-OH),2.27(t,2H,J=6.8Hz,N(CH2)3-CH 2-CN),1.90-1.94(m,2H,NCH2-CH 2-(CH2)2CN),1.57-1.51(m,2H,N(CH2)2-CH 2-CH2CN),1.47(t,3H,J=7.3Hz,-CH2-CH 3).13C NMR(150MHz,CDCl3)δ171.9×2,136.1×2,131.5×2,127.9,126.7,126.3,126.0,122.7,122.6,122.5,122.3,120.5,120.2,119.2,109.7,109.5,106.2,105.6,61.8,45.8,41.6,29.0,22.8,17.0,15.2.ESI-MS m/z489.1[M+Na]+.
化合物7的制备
i)1-氰甲基-3-吲哚乙酸(7a)的制备
氩气保护下,100mL三口瓶中,加氢化钠(1028mg,25.7mmol,质量分数60%,分散在石蜡中)于40mL DMF中,在-5℃搅拌悬浮,加入10mL DMF溶解的3-吲哚乙酸(900mg,5.14mmol),搅拌30min后,滴加10mL DMF溶解的溴乙腈(1.03mL,15.4mmol),缓慢升至室温,反应过夜后,降至0℃以下,滴加10mL甲醇,再加适量水至得亮黄色溶液,用30mL***萃取除去石蜡油,水层用6N盐酸酸化至弱酸性,乙酸乙酯萃取(100mL×3次),合并乙酸乙酯层,无水Na2SO4干燥,真空蒸干,后经Sephadex LH-20凝胶柱色谱分离、甲醇洗脱得白色晶体(7a)314mg,收率29%。1H NMR(600MHz,DMSO-d6)δ12.33(s,1H,-CO2 H),7.58(d,1H,J=8.3Hz,Ar-H),7.56(d,1H,J=8.2Hz,Ar-H),7.36(s,1H,Ar-H),7.26(dt,1H,J=7.8Hz,1.0Hz,Ar-H),7.13(dt,1H,J=7.4Hz,0.9Hz,Ar-H),5.52(s,2H,-CH2 -CN),3.69(s,2H,-CH2 -CO2H).13C NMR(150MHz,DMSO-d6)δ173.4,136.3,128.7,127.6,122.8,120.4,119.9,117.2,110.3,110.2,34.2,31.1.
ii)2,3-二(1-氰甲基-3-吲哚)马来酸酐(7b)制备
按照化合物1c的制备方法,由化合物1b(226mg,1.45mmol)、草酰氯(274mg,2.18mmol)、化合物7a(310mg,1.45mmol)和三乙胺(293mg,2.9mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末(7b)235mg,收率40%。1H NMR(600MHz,DMSO-d6)δ8.05(s,2H,Ar-H),7.64(d,2H,J=8.3Hz,Ar-H),7.22(dt,2H,J=7.6Hz,1.2Hz,Ar-H),6.89(d,2H,J=7.9Hz,Ar-H),6.84(dt,2H,J=7.9Hz,0.8Hz,Ar-H),5.69(s,4H,-CH2 -CN).13C NMR(150MHz,DMSO-d6)δ166.0×2,135.6×2,132.9×2,128.6×2,125.5×2,123.2×2,121.6×2,121.1×2,115.9×2,110.4×2,105.8×2,34.2×2.ESI-MS m/z 407.2[M+H]+.
iii)2,3-二(1-氰甲基-3-吲哚)马来酰亚胺(7)的制备
按照化合物1的制备方法,由化合物7b(200mg,0.49mmol)、HMDS(4.1mL,19.6mmol)和MeOH(0.39mL,9.8mmol)制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得橙红色粉末(7)183mg,收率92%。1H NMR(600MHz,DMSO-d6)δ11.10(s,1H,imide-NH),7.94(s,2H,Ar-H),7.58(d,2H,J=8.3Hz,Ar-H),7.15(dt,2H,J=7.4Hz,1.1Hz,Ar-H),6.82(d,2H,J=8.0Hz,Ar-H),6.75(dt,2H,J=7.0Hz,0.8Hz,Ar-H),5.65(s,4H,-CH2 -CN).13C NMR(150MHz,DMSO-d6)δ172.4×2,135.5×2,131.8×2,127.8×2,126.1×2,122.7×2,121.3×2,120.6×2,116.1×2,110.1×2,106.5×2,34.1×2.ESI-MS m/z406.1[M+H]+.
化合物8的制备
i)1-氰丙基-3-吲哚乙酸(8a)的制备
按照化合物7a的制备方法,吲哚乙酸(1.4g,8mmol)、NaH(1.6g,40mmol,质量分数60%,分散在石蜡中)和溴丁腈(2.4mL,24mmol)为原料制得,经硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得白色晶体(8a)541mg,收率30%。1H NMR(600MHz,DMSO-d6)δ7.54(d,1H,J=7.3Hz,Ar-H),7.42(d,1H,J=7.7Hz,Ar-H),7.22(s,1H,Ar-H),7.12(t,1H,J=7.3Hz,Ar-H),7.00(1H,t,J=7.3Hz,Ar-H),4.17(t,2H,J=6.9Hz,N-CH2 -(CH2)2CN),3.53(s,2H,-CH2 -CO2H),2.43(t,2H,J=6.9Hz,N(CH2)2-CH2 -CN),2.03-2.01(2H,m,NCH2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ175.0,136.3,128.6,127.2,121.6,120.7,119.9,119.0,110.4,109.8,44.5,33.2,26.4,14.4.ESI-MS m/z 241.1[M–H]–.
ii)2,3-二(1-氰丙基-3-吲哚)马来酸酐(8b)的制备
按照化合物1c的制备方法,化合物3b(452mg,2.46mmol)、草酰氯(469mg,3.69mmol)、化合物8a(595mg,2.46mmol)和三乙胺(497mg,4.92mmol)为原料制得,经硅胶柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得红色粉末(8b)500mg,收率44%1H NMR(600MHz,DMSO-d6)δ7.88(s,2H,Ar-H),7.57(d,2H,J=8.4Hz,Ar-H),7.14(dt,2H,J=7.8Hz,0.9Hz,Ar-H),6.95(d,2H,J=8.4Hz,Ar-H),6.81(dt,2H,J=7.8Hz,0.9Hz,Ar-H),4.30(t,4H,J=6.9Hz,N-CH2 -(CH2)2CN),2.42(t,4H,J=7.3Hz,N(CH2)2-CH2 -CN),2.05-2.02(m,4H,NCH2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ166.8×2,136.6×2,133.7×2,128.5×2,125.7×2,123.0×2,122.2×2,120.8×2,120.4×2,111.1×2,105.1×2,45.3×2,26.1×2,14.3×2.ESI-MS m/z 463.2[M+H]+.
iii)2,3-二(1-氰丙基-3-吲哚)马来酰亚胺(8)的制备
按照化合物1的制备方法,由化合物8b(375mg,0.81mmol)、HMDS(6.8mL,32.5mmol)和MeOH(0.66mL,16.2mmol)制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得橙红色粉末(8)341mg,收率91%。1H NMR(600MHz,DMSO-d6)δ10.98(s,1H,imide-NH),7.76(s,2H,Ar-H),7.50(d,2H,J=8.2Hz,Ar-H),7.08(dt,2H,J=7.8Hz,0.9Hz,Ar-H),6.90(d,2H,J=8.2Hz,Ar-H),6.73(dt,2H,J=7.4Hz,0.9Hz,Ar-H),4.28(t,4H,J=6.9Hz,N-CH2 -(CH2)2CN),2.38(4H,t,J=7.3Hz,N(CH2)2-CH2 -CN),2.04-2.02(m,4H,NCH2-CH2 -CH2CN).13CNMR(150MHz,DMSO-d6)δ173.3×2,136.4×2,132.5×2,128.2×2,126.2×2,122.5×2,121.9×2,120.5×2,120.2×2,110.7×2,105.8×2,45.0×2,26.1×2,14.3×2.HR-ESIMSm/z 460.1769[M–H]–(calcd.for C28H22N5O2,460.1774).
化合物9的制备
i)1-氰丁基吲哚-3-乙酸(9a)的制备
按照化合物7a的制备方法,吲哚乙酸(1.4g,8mmol),NaH(1.6g,40mmol,60%分散在石蜡中)和溴戊腈(2mL,16mmol)为原料制得,经硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得白色晶体(9a)1.69g,收率83%。1H NMR(600MHz,DMSO-d6)δ12.30(s,1H,-CO2 H),7.59(d,1H,J=7.8Hz,Ar-H),7.45(d,1H,J=8.2Hz,Ar-H),7.27(s,1H,Ar-H),7.18(dt,1H,J=7.8Hz,0.9Hz,Ar-H),7.08(dt,1H,J=7.8Hz,0.9Hz,Ar-H),4.13(t,2H,J=6.9Hz,N-CH 2-(CH2)3CN),3.72(s,2H,-CH2 -CO2H),2.44(t,2H,J=6.9Hz,N(CH2)3-CH2 -CN),1.81-1.79(m,2H,NCH2-CH 2-(CH2)2CN),1.51-1.48(m,2H,N(CH2)2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ173.7,136.4,128.2,127.7,121.8,121.0,119.6,119.2,110.2,107.9,45.1,31.4,29.6,22.8,16.3.ESI-MS m/z 255.1[M–H]–.
ii)2,3-二(1-氰丁基-3-吲哚)马来酸酐(9b)的制备
按照化合物1c的制备方法,化合物5b(1140mg,5.76mmol)、草酰氯(1097mg,8.64mmol)、化合物9a(1470mg,5.76mmol)和三乙胺(1163mg,11.5mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得产物红色粉末(9b)900mg,收率32%。1HNMR(600MHz,DMSO-d6)δ7.93(s,2H,Ar-H),7.55(d,2H,J=8.3Hz,Ar-H),7.09(t,2H,J=7.3Hz,Ar-H),6.83(d,2H,J=8.3Hz,Ar-H),6.73(2H,t,J=7.4Hz,Ar-H),4.32(t,4H,J=6.9Hz,N-CH 2-(CH2)3CN),2.52(t,4H,J=7.3Hz,N(CH2)3-CH2 -CN),1.85-1.83(m,4H,NCH2-CH 2-(CH2)2CN),1.51-1.48(m,4H,N(CH2)2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ166.9×2,136.4×2,133.6×2,128.3×2,126.0×2,122.8×2,121.9×2,121.0×2,120.6×2,111.2×2,104.9×2,45.7×2,29.3×2,22.7×2,16.4×2.ESI-MS m/z 491.2[M+H]+.
iii)2,3-二(1-氰丁基-3-吲哚)马来酰亚胺(9)的制备
按照化合物1的制备方法,由化合物9b(470mg,0.96mmol)、HMDS(8.1mL,38.4mmol)和MeOH(0.77mL,19.2mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=3:2(v/v)洗脱得橙红色粉末(9)426mg,收率90%。1H NMR(600MHz,DMSO-d6)δ10.93(s,1H,imide-NH),7.80(s,2H,Ar-H),7.48(d,2H,J=8.2Hz,Ar-H),7.02(dt,2H,J=7.8Hz,1.0Hz,Ar-H),6.77(d,2H,J=8.2Hz,Ar-H),6.63(dt,2H,J=7.4Hz,0.9Hz,Ar-H),4.29(t,4H,J=6.9Hz,N-CH 2-(CH2)3CN),2.51(t,4H,J=7.3Hz,N(CH2)3-CH2 -CN),1.83-1.81(m,4H,NCH2-CH 2-(CH2)2CN),1.50-1.48(m,4H,N(CH2)2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ173.4×2,136.3×2,132.4×2,128.0×2,126.5×2,122.3×2,121.6×2,121.0×2,120.0×2,110.8×2,105.6×2,45.5×2,29.3×2,22.7×2,16.3×2.ESI-MS m/z 488.2[M–H]–.
化合物10的制备
在25mL单口瓶中将化合物1(50mg,0.127mmol)用10mL乙酸:浓盐酸=3:1(v/v)的混合液溶解,120℃回流30min后,降至室温,加水和乙酸乙酯萃取,有机层蒸干,硅胶柱色谱分离、二氯甲烷洗脱得红色固体2-(1-乙基-3-吲哚)-3-(1-羧甲基-3-吲哚)马来酰亚胺(10)50mg,收率95%。1H NMR(600MHz,DMSO-d6)δ10.97(brs,1H,imide-NH),7.88(s,1H,Ar-H),7.66(s,1H,Ar-H),7.45(d,1H,J=8.2Hz,Ar-H),7.32(d,1H,J=8.2Hz,Ar-H),7.04(t,1H,J=7.1Hz,Ar-H),7.02(d,1H,J=8.2Hz,Ar-H),6.97(t,1H,J=8.2Hz,Ar-H),6.72(t,1H,J=7.1Hz,Ar-H),6.60(d,1H,J=7.7Hz,Ar-H),6.58(t,1H,J=7.7Hz,Ar-H),4.86(s,2H,-CH2 -CO2H),4.21(q,2H,J=7.2Hz,-CH2 -CH3),1.27(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.6,173.5,172.2,137.2,135.9,134.2,131.7,128.1,127.2,126.9,126.0,122.2,122.1,122.0,121.7,120.1,119.9,111.0,110.5,105.6,105.2,49.9,41.1,15.8.ESI-MS m/z 412.0[M–H]–.
化合物11的制备
在50mL两口瓶中,用10mL DMF悬浮NaH(68mg,1.7mmol,质量分数60%,分散在石蜡中),-5℃搅拌条件下加入10mL DMF溶解的化合物1(200mg,0.56mmol),低温反应30min后,缓慢滴加溴代乙腈(114μL,1.7mmol),低温反应30min。滴加饱和NH4Cl溶液终止反应,乙酸乙酯萃取,有机层浓缩,凝胶柱色谱分离、甲醇洗脱得红色固体粉末N-氰甲基-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(11)161mg,收率66%。1H NMR(600MHz,DMSO-d6)δ7.93(s,1H,Ar-H),7.89(s,1H,Ar-H),7.60(d,1H,J=8.2Hz,Ar-H),7.50(d,1H,J=8.2Hz,Ar-H),7.18(t,1H,J=7.1Hz,Ar-H),7.08(t,1H,J=7.2Hz,Ar-H),6.86(d,1H,J=8.2Hz,Ar-H),6.85(d,1H,J=8.3Hz,Ar-H),6.79(t,1H,J=7.1Hz,Ar-H),6.72(t,1H,J=7.7Hz,Ar-H),5.66(s,2H,N-CH2 -CN),4.75(s,2H,N-CH2 -CN),4.27(q,2H,J=7.1Hz,-CH2 -CH3),1.33(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ170.3,170.2,136.2×2,133.0,132.3,129.5,126.4,126.3,125.9,123.4,122.6,122.1,121.9,121.2,120.5,116.7,116.3,110.9,110.8,107.1,105.1,41.4,34.7,26.4,15.7.ESI-MS m/z 434.1[M+H]+.
化合物12的制备
按照化合物10的制备方法,以化合物11(50mg,0.12mmol)为原料制备,硅胶柱色谱分离、二氯甲烷:甲醇=5:1(v/v)洗脱得红色固体N-羧甲基-2-(1-乙基-3-吲哚)-3-(1-羧甲基-3-吲哚)马来酰亚胺(12)48mg,收率85%。1H NMR(600MHz,DMSO-d6)δ7.93(s,1H,Ar-H),7.80(s,1H,Ar-H),7.47(d,1H,J=8.3Hz,Ar-H),7.38(d,1H,J=8.2Hz,Ar-H),7.05(t,1H,J=7.7Hz,Ar-H),7.02(t,1H,J=7.1Hz,Ar-H),6.96(d,1H,J=7.7Hz,Ar-H),6.72(t,1H,J=7.7Hz,Ar-H),6.70(d,1H,J=8.2Hz,Ar-H),6.65(t,1H,J=7.1Hz,Ar-H),5.13(s,2H,N-CH2 -CO2H),4.29(s,2H,-CH2 -CO2H),4.25(q,2H,J=7.1Hz,-CH2 -CH3),1.32(t,3H,J=7.1Hz,-CH2-CH 3).13C NMR(150MHz,DMSO-d6)δ171.6,171.5,170.5,170.0,137.2,136.0,134.0,132.2,127.6,126.9,126.6,126.1,122.5,122.4,122.1,121.7,120.4,120.3,110.9,110.7,105.8,105.4,48.1,41.3,39.9,15.7.ESI-MS m/z 470.0[M–H]–.
化合物13的制备
i)1,2,3,4,6-D-吡喃葡萄糖五乙酸酯(13a)的制备
在100mL单口瓶中,加葡萄糖(2g,11.1mmol)、无水乙酸钠(2.5g,30.5mmol)、乙酸酐12.5mL,110℃回流,趁热将反应液倒入约100g碎冰中,搅拌产生大量白色固体,冰融化后抽滤,滤饼用无水乙醇重结晶,得白色粉末(13a)4.1g,收率95%。ESI-MS m/z 391.1[M+H]+.
ii)2,3,4,6-O-四乙酰-D-吡喃葡萄糖(13b)的制备
在N2保护下,在50mL两口瓶中,用10mL无水THF溶解13a(525mg,1.35mmol),–5℃下滴加苄胺(0.22mL,2.02mmol),缓慢升至室温,反应过夜,TLC检测反应完全,真空蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得白色固体(13b)441mg,收率94%。ESI-MSm/z 349.2[M+H]+.iii)2,3,4,6-O-四乙酰-D-吡喃葡萄糖三氯乙酸亚胺酯(13c)的制备
在两口瓶中,N2保护下,用5mL CH2Cl2溶解13b(390mg,1.12mmol),–5℃下滴加三氯乙腈(1.35mL,13.45mmol),滴加催化量的DBU,反应液由微黄色变为浅黄色,反应30min后,真空蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得乳白色固体(13c)381mg,收率70%。ESI-MS m/z 492.0[M+H]+.
iv)O-2-(1-乙基-3-吲哚)-3-(1-氰乙基-3-吲哚)马来酰亚胺-2,3,4,6-O-四乙酰-α-D-吡喃葡萄糖糖苷(13d)
将化合物13c(10mg,22.8μmol)和化合物4(7.5mg,16.5μmol)用气泵抽干后加到15mL两口瓶中,在干燥器中抽3h。分子筛用马弗炉烘干后粉碎,粉末用酒精喷灯烧30min,气泵抽冷后加约200mg至反应瓶中,加入5mL干燥的CH2Cl2,气泵换N2三次,降至–20℃下反应20min,滴加2μL BF3·Et2O,立即由红色变为紫色,继而回复红色,升至室温反应10h,反应完全。降至–5℃,加10mg NaHCO3终止反应,抽滤,蒸干溶剂,凝胶柱色谱分离、二氯甲烷:甲醇=1:1(v/v)洗脱得红色固体(13d)16.7mg,收率95%。1H NMR(600MHz,CDCl3)δ7.79(s,1H,Ar-H),7.61(s,1H,Ar-H),7.31(d,1H,J=8.2Hz,Ar-H),7.30(d,1H,J=8.2Hz,Ar-H),7.17(t,1H,J=8.3Hz,Ar-H),7.11(t,1H,J=8.2Hz,Ar-H),7.08(d,1H,J=8.2Hz,Ar-H),6.86(d,1H,J=7.3Hz,Ar-H),6.85(t,1H,J=7.3Hz,Ar-H),6.77(t,1H,J=7.3Hz,Ar-H),5.36/5.28(d,2H,J=11.5Hz,N-CH2 -O),5.19(t,1H,J=10.0Hz,Glc-C3-H),5.10(t,1H,J=10.0Hz,Glc-C2-H),5.00(t,1H,J=10.0Hz,Glc-C4-H),4.86(d,1H,J=8.2Hz,Glc-C1-H),4.46(t,2H,J=7.3Hz,N-CH2 -CH2CN),4.21(q,2H,J=7.3Hz,N-CH2 -CH3),4.18/4.03(dd,2H,J=12.4Hz,2.8Hz,Glc-C6-H2 ),3.70(dt,1H,J=10.1Hz,3.7Hz,Glc-C5-H),2.82(t,2H,J=7.3Hz,NCH2-CH2 -CN),1.94(s,12H,4-COCH3 ),1.49(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,CDCl3)δ171.4,171.3,170.8,170.4,169.5,169.4,136.2,135.5,131.9,130.9,129.0,126.5,125.9,125.8,123.2,122.8,122.5,122.4,121.0,120.5,116.8,109.8,109.0,107.3,105.5,100.0,73.0,72.1,71.2,68.1,66.2,61.6,42.4,41.6,20.8,20.7,20.6,20.6,19.0,15.3.ESI-MS m/z 791.4[M+Na]+.
v)O-2-(1-乙基-3-吲哚)-3-(1-氰乙基-3-吲哚)马来酰亚胺-α-D-吡喃葡萄糖糖苷(13)的制备
在单口瓶中,将样品13d用1mL CH2Cl2溶解,加4mL无水甲醇,0℃搅拌下滴加NaOMe/MeOH,至pH 9~10,升至室温反应30min,TLC检测无原料剩余,0℃下加饱和NH4Cl溶液终止反应。乙酸乙酯萃取,蒸干,凝胶柱色谱分离、甲醇洗脱得红色固体(13)13mg,收率100%。1HNMR(600MHz,DMSO-d6)δ7.94(s,1H,Ar-H),7.87(s,1H,Ar-H),7.60(d,1H,J=8.2Hz,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.06(t,1H,J=7.8Hz,Ar-H),7.04(t,1H,J=7.3Hz,Ar-H),6.82(d,1H,J=7.8Hz,Ar-H),6.78(d,1H,J=8.3Hz,Ar-H),6.68(t,2H,J=7.8Hz,Ar-H),5.20/5.14(d,2H,J=11.0Hz,N-CH2 -O),5.10(d,1H,J=5.5Hz,Glc-C1-H),4.99(d,1H,J=3.7Hz,Glc-C2-OH),4.92(d,1H,J=3.7Hz,Glc-C3-OH),4.60(t,2H,J=6.7Hz,N-CH2 -CH2CN),4.47(t,1H,J=6.0Hz,Glc-C6-OH),4.41(d,1H,J=8.2Hz,Glc-C4-OH),4.27(q,2H,J=7.3Hz,N-CH2 -CH3),3.62(m,1H,Glc-C2-H),3.48(m,1H,Glc-C3-H),3.11(m,2H,Glc-C6-H2 ),3.09(m,1H,Glc-C4-H),3.04(t,2H,J=6.7Hz,NCH2-CH2 -CN),2.95(m,1H,Glc-C5-H),1.34(t,3H,J=7.3Hz,N-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.5,171.4,136.1,136.0,132.5,132.4,128.4,126.7,126.4,126.3,122.6,122.4,122.0,121.8,120.5,120.4,119.0,110.9,110.7,106.2,105.3,103.0,77.6,77.3,73.8,70.2,66.2,61.4,41.9,41.3,19.1,15.8.HR-ESIMS m/z 623.2139[M+Na]+(C33H35N4O8Na,623.2118).
化合物14的制备
将1c(20mg,50.6μmol)用2mL DMF溶解,搅拌下加入0.5mL乙二胺,室温反应过夜,加入适量水,乙酸乙酯萃取,有机层蒸干,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得橙红色粉末N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(14)22mg,收率99%。1H NMR(600MHz,DMSO-d6)δ7.89(s,1H,Ar-H),7.84(s,1H,Ar-H),7.60(d,1H,J=8.2Hz,Ar-H),7.49(d,1H,J=8.2Hz,Ar-H),7.17(t,1H,J=7.8Hz,Ar-H),7.07(t,1H,J=7.4Hz,Ar-H),6.88(d,1H,J=7.8Hz,Ar-H),6.86(d,1H,J=7.6Hz,Ar-H),6.78(t,1H,J=7.4Hz,Ar-H),6.71(t,1H,J=7.6Hz,Ar-H),5.67(s,2H,-CH2 -CN),4.26(q,2H,J=7.3Hz,N-CH2 -CH3),3.71(t,2H,J=6.2Hz,N-CH2 -CH2-NH2),2.94(t,2H,J=6.2Hz,N-CH2-CH2 -NH2),1.32(t,3H,J=7.3Hz,N-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.9×2,136.0,135.9,132.4,132.0,129.0,126.4,126.2,125.7,123.1,122.4,122.1,121.9,121.0,120.2,116.7,110.7,110.6,107.3,105.2,41.2,40.0,39.0,34.6,15.7.ESI-MS m/z 438.1[M+H]+.
化合物15的制备
按照化合物14的制备方法,以化合物2c(60mg,146.7μmol)为原料制备,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得红色晶体N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(1-氰乙基-3-吲哚)马来酰亚胺(15)74.5mg,收率100%。1H NMR(600MHz,DMSO-d6)δ7.89(s,1H,Ar-H),7.82(s,1H,Ar-H),7.60(d,1H,J=8.3Hz,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.07(t,1H,J=7.8Hz,Ar-H),7.05(t,1H,J=7.3Hz,Ar-H),6.87(d,1H,J=8.0Hz,Ar-H),6.83(d,1H,J=8.0Hz,Ar-H),6.70(t,1H,J=7.3Hz,Ar-H),6.69(t,1H,J=7.8Hz,Ar-H),4.59(t,2H,J=6.4Hz-CH2 -CH2-CN),4.27(q,2H,J=7.2Hz,N-CH2 -CH3),3.76(t,2H,J=6.1Hz,N-CH2 -CH2-NH2),3.03(t,2H,J=6.1Hz,N-CH2-CH2 -NH2),3.00(t,2H,J=6.4Hz,-CH2 -CN),1.33(t,3H,J=7.2Hz,N-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.4,171.3,135.5,135.4,131.5,131.4,127.7,126.4,125.9,125.8,121.9,121.7,121.5,121.2,119.7,119.6,118.3,110.2,110.0,105.7,104.9,41.3,40.6,38.5,37.4,18.4,15.1.ESI-MS m/z452.3[M+H]+.
化合物16的制备
将化合物15(20mg,0.044mmol)溶解于4N盐酸/乙酸乙酯溶液中,室温搅拌6h,蒸干溶剂,冷冻干燥5h后,无水乙醇/石油醚(v/v,5:1)重结晶得深红色晶体N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(1-氰乙基-3-吲哚)马来酰亚胺盐酸盐(16)17mg,收率79.2%。1H NMR(600MHz,DMSO-d6)δ8.15(brs,3H,-NH3 +),7.92(s,1H,Ar-H),7.85(s,1H,Ar-H),7.65(d,1H,J=8.0Hz,Ar-H),7.52(d,1H,J=8.0Hz,Ar-H),7.07(t,1H,J=7.8Hz,Ar-H),7.15(t,1H,J=7.6Hz,Ar-H),6.89(d,1H,J=8.0Hz,Ar-H),6.81(d,1H,J=8.0Hz,Ar-H),6.72(t,1H,J=7.3Hz,Ar-H),6.70(t,1H,J=7.8Hz,Ar-H),4.58(t,2H,J=6.4Hz N-CH2 -CH2CN),4.27(q,2H,J=7.2Hz,N-CH2 -CH3),3.96(t,2H,J=6.2Hz,N-CH2 -CH2-NH3 +),3.73(t,2H,J=6.2Hz,N-CH2-CH2 -NH3 +),3.01(t,2H,J=6.4Hz,NCH2-CH2 -CN),1.32(t,3H,J=7.2Hz,N-CH2-CH3 ).13CNMR(150MHz,DMSO-d6)δ171.4,171.2,135.6,135.4,131.5,131.3,127.7,127.4,125.9,125.8,121.9,121.6,121.5,121.2,119.7,119.5,118.3,110.2,110.1,105.7,104.7,45.3,42.6,38.5,37.4,18.4,15.1.ESI-MS m/z 452.2[M–Cl]+.
化合物17的制备
按照化合物14的制备方法,以化合物3c(24mg,56.7μmol)和乙二胺为原料制备,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得深红色固体N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(1-氰丙基-3-吲哚)马来酰亚胺(17)26mg,收率98%。1H NMR(600MHz,DMSO-d6)δ7.79(s,1H,Ar-H),7.76(s,1H,Ar-H),7.53(d,1H,J=8.3Hz,Ar-H),7.50(d,1H,J=8.3Hz,Ar-H),7.09(t,1H,J=8.5Hz,Ar-H),7.07(t,1H,J=8.5Hz,Ar-H),6.90(t,2H,J=7.9Hz,Ar-H),6.75(d,1H,J=7.4Hz,Ar-H),6.72(d,1H,J=7.4Hz,Ar-H),4.31(t,2H,J=6.7Hz,N-CH2 -(CH2)2CN),4.25(q,2H,J=7.2Hz,N-CH2 -CH3),3.76(t,2H,J=5.7Hz,N-CH2 -CH2-NH2),3.01(t,2H,J=5.7Hz,NCH2-CH2 -NH2),2.41(t,2H,J=7.2Hz,N(CH2)2-CH2 -CN),2.02(m,2H,NCH2-CH2 -CH2CN),1.31(t,3H,J=7.2Hz,N-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.5×2,135.9,135.6,131.9,131.8,127.4,126.4,125.6×2,122.0,121.8,121.6×2,120.0,119.8,119.6,110.3,110.2,105.4,104.9,44.5,40.7,38.5,37.2,25.6,15.2,13.7.HR-ESIMS m/z 466.2266[M+H]+(calcd.for C28H28N5O2,466.2243).
化合物18的制备
按照化合物14的制备方法,以化合物4c(24mg,54.9μmol)为原料制备,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得红色固体N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(1-氰丁基-3-吲哚)马来酰亚胺(18)30mg,收率100%。1H NMR(600MHz,DMSO-d6)δ7.82(s,1H,Ar-H),7.80(s,1H,Ar-H),7.51(d,1H,J=8.4Hz,Ar-H),7.47(d,1H,J=8.4Hz,Ar-H),7.05(t,1H,J=7.5Hz,Ar-H),7.04(t,1H,J=7.4Hz,Ar-H),6.83(d,1H,J=8.5Hz,Ar-H),6.82(d,1H,J=8.7Hz,Ar-H),6.68(t,1H,J=7.5Hz,Ar-H),6.66(t,1H,J=7.4Hz,Ar-H),4.30(t,2H,J=6.8Hz,N-CH2 -(CH2)3CN),4.26(q,2H,J=7.2Hz,N-CH2 -CH3),3.60(t,2H,J=6.4Hz,N-CH2 -CH2-NH2),2.82(t,2H,J=6.4Hz,N-CH2-CH2 -NH2),2.51(t,2H,J=7.2Hz,N(CH2)3-CH2 -CN),1.82(m,2H,NCH2-CH2 -(CH2)2CN),1.46(m,2H,N(CH2)2-CH2 -CH2CN),1.33(t,3H,J=7.2Hz,N-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ172.2×2,136.2,136.0,132.4,132.0,127.9,127.3,126.8,126.3,122.3,122.2,121.8,121.0,120.0,119.9,114.3,110.7,110.6,105.6,105.4,45.4,41.2,40.8,40.0,29.2,22.6,16.3,15.7.ESI-MS m/z480.2[M+H]+.
化合物19的制备
i)N-叔丁基氧羰基甘氨酸(19a)的制备
在15mL单口瓶中,以4mL 10%Na2CO3水溶液溶解甘氨酸(1.0g,13mmol),–5℃下滴加以2mL乙腈溶解的Boc酸酐(2.84g,13mmol),滴毕升至室温,搅拌过夜,过滤,滤液以石油醚萃取,弃掉石油醚层,水层用盐酸调至酸性,乙酯萃取,干燥,真空蒸干,硅胶柱色谱分离、二氯甲烷:甲醇=30:1(v/v)洗脱得白色针状晶体(19a)0.8g,收率35%。ESI-MS m/z 389.3[2M+K]+.
ii)N-(N-叔丁基氧羰基甘氨酰乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(19b)的制备
在25mL两口瓶中,以2mL CH2Cl2溶解化合物19a(26mg,0.15mmol)、化合物14(50mg,0.11mmol)和DMAP(4mg,0.03mmol),加入0.5mL CH2Cl2溶解的DCC(35mg,0.17mmol),室温反应过夜,TLC检测反应完全,抽滤,滤液蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=1:3(v/v)洗脱得橙红色固体(19b)47mg,收率69%。1H NMR(600MHz,DMSO-d6)δ8.05(t,1H,J=5.5Hz,-NH),7.87(s,1H,,Ar-H),7.86(s,1H,Ar-H),7.59(d,1H,J=8.2Hz,Ar-H),7.50(d,1H,J=8.2Hz,Ar-H),7.17(dt,1H,J=7.2Hz,1.1Hz,Ar-H),7.07(dt,1H,J=7.2Hz,1.1Hz,Ar-H),6.90(t,1H,J=5.9Hz,-NH),6.86(d,1H,J=8.2Hz,Ar-H),6.85(d,1H,J=8.2Hz,Ar-H),6.79(t,1H,J=7.7Hz,Ar-H),6.70(t,1H,J=7.7Hz,Ar-H),5.64(s,2H,-CH2 -CN),4.27(q,2H,J=7.1Hz,N-CH2 -CH3),3.65(t,2H,J=6.1Hz,N-CH2 -CH2-NH),3.51(m,2H,N-CH2-CH2 -NH),3.35('q'like,J=5.5Hz,6.1Hz,-CO-CH2 -NH-),1.35(s,9H,-C(CH3)3 ),1.33(t,3H,J=7.1Hz,N-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.9,171.8,171.0,169.0,136.1×2,132.5,132.0,129.1,126.6,126.3,125.6,123.3,122.5,122.2,121.9,121.1,120.3,116.7,110.8,110.6,107.5,105.3,78.6,60.4,41.3,38.1,37.8,34.6,28.7,15.8.ESI-MSm/z 495.3[M+H]+.
iii)N-甘氨酰乙基-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(19)的制备
以8mL甲苯溶解化合物19b(30mg,0.05mmol),加入适量100~200目硅胶,N2保护下110℃冷凝水回流3.5h,冷却至室温,减压柱抽滤,有机层蒸干,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得红色固体(19)23.5mg,收率95%。1H NMR(600MHz,DMSO-d6)δ7.87(s,1H,Ar-H),7.82(s,1H,Ar-H),7.59(d,1H,J=8.3Hz,Ar-H),7.49(d,1H,J=8.3Hz,Ar-H),7.17(dt,1H,J=7.7Hz,1.1Hz,Ar-H),7.07(dt,1H,J=7.7Hz,1.1Hz,Ar-H),6.86(d,1H,J=7.7Hz,Ar-H),6.84(d,1H,J=7.1Hz,Ar-H),6.79(t,1H,J=7.7Hz,Ar-H),6.71(t,1H,J=7.7Hz,Ar-H),5.65(s,2H,-CH2 -CN),4.25(q,2H,J=7.1Hz,N-CH2 -CH3),3.66(t,2H,J=5.5Hz,N-CH2 -CH2NH),3.50-3.60(brs.2H,-NH2),3.49(t,2H,J=5.5Hz,NCH2-CH2 -NH),3.23(s,2H,-CO-CH2 -NH2),3.15(s,1H,NCH2-CH2-NH),1.31(t,3H,J=7.1Hz,N-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.9×2,170.7,136.1×2,132.5,132.0,129.1,126.5,126.3,125.7,123.3,122.5,122.2,122.0,121.1,120.3,116.7,110.8,110.7,107.4,105.3,63.2,43.1,38.1,37.8,34.6,15.8.ESI-MS m/z 495.3[M+H]+.
化合物20的制备
i)N-叔丁基氧羰基-L-丙氨酸(20a)的制备
按照化合物19a的制备方法,以L-丙氨酸(1.3g,15mmol)为原料得白色针状晶体(20a)0.66g,收率23%。ESI-MS m/z 212.2[M+Na]+.)
ii)N-(N-叔丁基氧羰基-L-丙氨酰乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(20b)的制备
按照化合物19b的制备方法,以化合物20a(30mg,0.16mmol)、化合物14(50mg,0.11mmol)、DMAP(4mg,0.03mmol)和DCC(35mg,0.17mmol)为原料得橙红色固体(20b)45mg,收率65%。1H NMR(600MHz,DMSO-d6)δ8.05(t,1H,J=5.5Hz,-NH),7.85(s,1H,Ar-H),7.82(s,1H,Ar-H),7.58(d,1H,J=8.2Hz,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.16(dt,1H,J=7.1Hz,1.1Hz,Ar-H),7.06(dt,1H,J=7.1Hz,1.1Hz,Ar-H),6.85(d,2H,J=7.7Hz,Ar-H),6.77(t,1H,J=7.2Hz,Ar-H),6.70(t,1H,J=7.7Hz,Ar-H),5.64(s,2H,-CH2 -CN),4.24(q,2H,J=7.1Hz,N-CH2 -CH3),3.87(q,1H,J=8.3Hz,CH3-CH-NH-),3.62(m,2H,N-CH2 -CH2NH),3.47(m,1H,J=6.1Hz,NCH2-CH2 -NH-),3.26(m,1H,J=6.1Hz,NCH2-CH2 -NH-),1.32(s,9H,-C(CH3)3),1.30(t,3H,J=7.1Hz,N-CH2-CH3 ),1.13(d,3H,J=6.6Hz,-NH-CH-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.9,171.8,171.0,169.0,136.1×2,132.5,132.0,129.1,126.6,126.3,125.6,123.3,122.5,122.2,121.9,121.1,120.3,116.7,110.8,110.6,107.5,105.3,78.6,60.4,41.3,38.1,37.8,34.6,28.7,15.8.ESI-MS m/z 1219.2[2M+H]+.
iii)N-(L-丙氨酰乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(20)的制备
按照化合物19的制备方法,以化合物20b(30mg,0.049mmol)为原料制得产物(20)23.4mg,收率94%。1H NMR(600MHz,DMSO-d6)δ8.41(t,1H,J=5.5Hz,-NH),7.84(s,1H,Ar-H),7.67(s,1H,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.33(d,1H,J=8.3Hz,Ar-H),7.06(t,2H,J=7.1Hz,Ar-H),7.03(d,1H,J=8.3Hz,Ar-H),6.74(t,1H,J=7.1Hz,Ar-H),6.65(dd,2H,J=7.1Hz,8.3Hz,Ar-H),4.90(s,2H,-CH2 -CN),4.21(q,2H,J=7.1Hz,N-CH2 -CH3),3.66(m,2H,N-CH2 -CH2NH),3.49(m,2H,NCH2-CH2 -NH-),3.53(m,1H,J=6.6Hz,-CO-CH-NH2),1.28(t,3H,J=7.1Hz,-NCH2-CH3 ),1.18(d,3H,J=7.1Hz,H2NCH-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.4,172.1,169.4,137.1,136.0,134.1,131.9,127.2×2,126.7,126.0,122.4,122.3×2,122.2,121.9,120.2×2,110.8,110.6,105.6×2,49.7,49.1,41.2,38.0,37.4,19.7,15.7.ESI-MS m/z 509.2[M+H]+.
化合物21的制备
i)N,N-二叔丁基氧羰基-L-组氨酸(21a)的制备
按照化合物18a的制备方法,以组氨酸(1.0g,7mmol)为原料得白色针状晶体(21a)0.92g,收率37%。ESI-MS m/z 356.4[M+H]+.
ii)N-(N,N-二叔丁基氧羰基-L-组氨酰乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(21b)的制备
按照化合物18b的制备方法,以化合物21a(80.9mg,0.16mmol)、化合物14(50mg,0.11mmol)、DMAP(4.2mg,0.03mmol)和DCC(35mg,0.17mmol)为原料得产物(21b)35mg,收率45%。ESI-MS m/z 775.5[M+H]+.
iii)N-(L-组氨酰乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(21)的制备
按照化合物18的制备方法,以化合物21b(30mg,0.039mmol)为原料得产物(21)20.2mg,收率90%。1H NMR(600MHz,DMSO-d6)δ7.85(s,1H,Ar-H),7.80(s,1H,Ar-H),7.58(d,1H,J=8.2Hz,Ar-H),7.54(s,1H,imidazole-H),7.48(d,1H,J=8.2Hz,Ar-H),7.16(t,1H,J=7.1Hz,Ar-H),7.05(t,1H,J=7.1Hz,Ar-H),6.84(d,2H,J=8.2Hz,Ar-H),6.77(t,1H,J=6.6Hz,Ar-H),6.76(s,1H,imidazole-H),6.68(t,1H,J=7.7Hz,Ar-H),5.63(s,2H,-CH2 -CN),4.23(q,2H,J=7.1Hz,N-CH2 -CH3),3.65(m,2H,N-CH2 -CH2-NH),3.59(m,1H,NCH2-CH2 -NH-),3.53(m,1H,J=4.4Hz,-CO-CH-NH2),3.4(m,2H,imidazole-CH2 -CHNH2),1.29(t,3H,J=7.1Hz,NCH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.6,172.0,171.9,136.1,135.4,135.3,132.7,132.5,131.9,129.2,126.6,126.3,123.3,122.5,122.2,122.0,121.1,120.3,116.7,110.8,110.7,107.5,105.3,70.3,60.4,55.4,42.3,38.2,34.6,29.5,22.6,15.7.ESI-MS m/z575.3[M+H]+.
化合物22的制备
i)N-叔丁基氧羰基-L-色氨酸(22a)的制备
按照化合物18a的制备方法,以色氨酸(1.0g,5mmol)为原料得白色针状晶体(22a)1.3g,收率63.3%。ESI-MS m/z 305.3[M+H]+.
ii)N-(N-叔丁基氧羰基-L-色氨酰乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(22b)的制备
按照化合物18b的制备方法,以化合物22a(72mg,0.17mmol)、化合物13(50mg,0.11mmol)、DMAP(4.2mg,0.03mmol)和DCC(35mg,0.17mmol)为原料得化合物(22b)80mg,收率89%。ESI-MS m/z 724.2[M+H]+,746.2[M+Na]+.
iii)N-(L-色氨酰乙基)-2-(1-乙基-3-吲哚)-3-(1-氰甲基-3-吲哚)马来酰亚胺(22)的制备
按照化合物18的制备方法,以化合物22b(30mg,0.041mmol)为原料得产物(22)23.2mg,收率90%。1H NMR(600MHz,DMSO-d6)δ10.81(brs,1H,-NH),8.22(t,1H,J=6.1Hz,-NH),7.86(s,1H,Ar-H),7.81(s,1H,Ar-H),7.58(d,1H,J=8.3Hz,Ar-H),7.46(dd,2H,J=8.2Hz,7.7Hz,Ar-H),7.32(d,2H,J=8.2Hz,Ar-H),7.16(t,1H,J=8.8Hz,Ar-H),7.04–7.02(m,2H,Ar-H),6.88–6.86(m,3H,Ar-H),6.74(t,1H,J=7.7Hz,Ar-H),6.67(t,1H,J=7.2Hz,Ar-H),5.61(s,2H,-CH2 -CN),4.21(q,2H,J=7.1Hz,N-CH2 -CH3),3.67(m,2H,-N-CH2 -CH2-NH),3.43(m,2H,-CH-CH2 -indole),3.1(d,2H,J=3.8Hz,-CH-CH2 -indole),2.68–2.65(m,2H,NCH2-CH2 -NH-),1.26(t,3H,J=7.1Hz,NCH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ175.2,172.0,171.9,136.8,136.6,136.1,132.4,132.0,129.2,127.9,126.6,126.3,125.7,124.1,123.2,122.4,122.2,122.0,121.4,121.1,120.3,119.0,118.7,116.7,111.8,111.1,110.8,110.6,107.5,105.3,60.3,55.8,41.3,38.2,34.6,31.3,15.7.ESI-MS m/z624.4[M+H]+.
化合物23的制备
按照化合物16的制备方法,以化合物N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(930mg,2.34mmol)为原料得其盐酸盐23(893mg,收率88%)。1H NMR(500MHz,DMSO-d6)δ11.84(s,1H,indole-NH),8.19(brs,3H,-NH3 +),7.77(d,1H,J=2.5Hz,Ar-H),7.70(s,1H,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.39(d,1H,J=8.1Hz,Ar-H),7.05(t,1H,J=7.6Hz,Ar-H),6.99(dd,2H,J=7.1Hz,5.0Hz,Ar-H),6.74(t,1H,J=8.7Hz,Ar-H),6.73(t,1H,J=7.9Hz,Ar-H),6.63(t,1H,J=7.5Hz,Ar-H),4.25(q,2H,J=7.2Hz,-NCH2 -CH3),3.84(t,2H,J=5.7Hz,N-CH 2-CH2NH3 +),3.10(t,2H,J=5.7Hz,NCH2-CH2 -NH3 +),1.29(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.2,172.1,136.5,135.9,131.8,129.8,127.6,126.9,126.4,125.4,122.2,122.1,121.9,121.5,120.0,119.8,112.3,110.5,106.0,105.5,41.1,40.2,39.4,15.7.HR-ESIMS m/z 399.1826[M–Cl]+(calcd.for C24H23N4O2,399.1815).
化合物24的制备
i)1-苄基吲哚(24a)的制备
在100mL三口瓶中,将NaH(300mg,7.5mmol,质量分数60%,分散于石蜡中)用30mLDMF悬浮,–5℃下缓慢滴加10mL DMF溶解的吲哚(585mg,5mmol),升至室温反应30min再降至–5℃。滴加溴化苄(0.89mL,7.5mmol),滴加完毕,–5℃下搅拌反应30min,反应完全,加入10mL甲醇,后加100mL饱和氯化铵溶液,CH2Cl2萃取(100mL×3),合并有机层,无水硫酸钠干燥,真空蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=100:1(v/v)洗脱得乳白色固体(24a)1.02g,收率99%。1H NMR(600MHz,CDCl3)δ7.76(d,1H,J=7.7Hz,Ar-H),7.38–7.33(m,4H,Ar-H),7.27(dt,1H,J=6.8Hz,0.9Hz,Ar-H),7.22(dt,1H,J=6.9Hz,0.9Hz,Ar-H),7.20(t,1H,J=3.2Hz,Ar-H),7.18(d,2H,J=6.8Hz,Ar-H),6.65(dd,1H,J=3.2Hz,0.9Hz,Ar-H),5.37(s,2H,Ph-CH2-).13C NMR(150MHz,CDCl3)δ137.7,136.5,128.9,128.9,128.4,127.8,127.0,126.9,121.9,121.2,119.7×2,109.9,101.9,50.2.ESI-MS m/z 208.2[M+H]+.
ii)2-(1-乙基-3-吲哚)-3-(1-苄基-3-吲哚)马来酸酐(24b)的制备
按照化合物1c的制备方法,以化合物24a(356mg,1.72mmol)、草酰氯(328mg,2.58mmol)、化合物1a(349mg,1.72mmol)和Et3N(347mg,3.44mmol)为原料得红色固体(24b)299mg,收率39%。1H NMR(600MHz,DMSO-d6)δ8.02(s,1H,Ar-H),7.94(s.1H,Ar-H),7.53(d,1H,J=7.3Hz,Ar-H),7.44(d,1H,J=7.8Hz,Ar-H),7.33(t,2H,J=6.4Hz,Ar-H),7.27(t,1H,J=6.8Hz,Ar-H),7.20(d,2H,J=5.9Hz,Ar-H),7.11(t,1H,J=7.3Hz,Ar-H),7.05(t,1H,J=6.8Hz,Ar-H),6.91(d,1H,J=7.3Hz,Ar-H),6.88(d,1H,J=7.3Hz,Ar-H),6.75(t,1H,J=7.3Hz,Ar-H),6.73(t,1H,J=7.3Hz,Ar-H),5.52(s,2H,Ph-CH2-),4.29(q,2H,J=6.8Hz,-CH2 -CH3),1.34(t,3H,J=6.8Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ167.0,166.9,137.9,136.6,136.3,133.9,133.3,129.2×2,128.8,128.1,127.8,127.6×2,126.2,125.9,122.9,122.8,122.2,122.0,120.8,120.7,111.5,111.1,105.3,104.7,50.0,40.5,15.7.ESI-MS m/z447.2[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-3-吲哚)马来酰亚胺(24c)的制备
按照化合物1的制备方法,由化合物24b(260mg,0.58mmol)、HMDS(2.44mL,11.7mmol)和MeOH(0.23mL,5.8mmol)制备得红色粉末状固体(24c)248mg,收率96%。1H NMR(600MHz,DMSO-d6)δ10.95(s,1H,imide-NH),7.90(s,1H,Ar-H),7.83(s,1H,Ar-H),7.46(d,1H,J=8.2Hz,Ar-H),7.36(d,1H,J=8.2Hz,Ar-H),7.32(t,2H,J=7.6Hz,Ar-H),7.26(t,1H,J=7.3Hz,Ar-H),7.17(d,2H,J=7.3Hz,Ar-H),7.04(t,1H,J=7.8Hz,Ar-H),6.97(t,1H,J=7.3Hz,Ar-H),6.84(d,1H,J=8.2Hz,Ar-H),6.82(d,1H,J=7.7Hz,Ar-H),6.65(t,1H,J=7.8Hz,Ar-H),6.64(t,1H,J=7.4Hz,Ar-H),5.49(s,2H,Ph-CH2-),4.26(q,2H,J=7.3Hz,-CH2 -CH3),1.34(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.4×2,138.2,136.4,136.0,132.8,132.0,129.1×2,128.5,128.0,127.5,127.4×2,126.7,126.4,122.4,122.2,121.8,121.7,120.2,120.1,111.1,110.7,106.0,105.4,49.9,41.3,15.8.ESI-MS m/z446.2[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(24d)的制备
将化合物24c(100mg,0.225mmol)以DMSO(0.85mL)溶解,搅拌条件下,滴加1M的t-BuOK/THF溶液(8.4mL,8.4mmol),滴加完毕,向反应液中通入O2约30min,加饱和氯化铵溶液终止反应,乙酸乙酯萃取(100mL×3次),合并有有机层,并用无水Na2SO4干燥,真空蒸干。硅胶柱色谱分离、二氯甲烷:乙酸乙酯=6:1(v/v)洗脱得红色粉末(24d)70.5mg,收率89%。1HNMR(600MHz,DMSO-d6)δ11.66(s,1H,indole-NH),10.90(s,1H,imido-NH),7.76(s,1H,Ar-H),7.72(s,1H,Ar-H),7.46(d,1H,J=8.2Hz,Ar-H),7.37(d,1H,J=8.1Hz,Ar-H),7.04(t,1H,J=7.6Hz,Ar-H),6.98(t,1H,J=7.5Hz,Ar-H),6.90(d,1H,J=8.0Hz,Ar-H),6.74(d,1H,J=8.1Hz,Ar-H),6.69(t,1H,J=7.5Hz,Ar-H),6.62(t,1H,J=7.5Hz,Ar-H),4.24(q,2H,J=7.2Hz,-CH2 -CH3),1.31(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.6,173.5,136.6,136.0,131.9,129.8,128.4,127.7,126.6,125.7,122.2,122.1,121.9,121.6,120.1,119.9,112.3,110.6,106.1,105.5,41.2,15.8.ESI-MS m/z 356.1[M+H]+.
v)2-(1-乙基吲哚)-3-(3-吲哚)马来酸酐(24e)的制备
在50mL单口瓶中,用20mL 10%的KOH水溶液悬浮化合物24d(50mg,0.14mmol),110℃下回流40min后冷却至室温,滴加2N盐酸酸化,乙酸乙酯萃取,合并有有机层,并用无水硫酸钠干燥,真空浓缩,硅胶柱色谱分离、二氯甲烷洗脱得红色固体(24e)43mg,收率86%。1HNMR(600MHz,DMSO-d6)δ11.96(s,1H,indole-NH),7.89(d,1H,J=2.8Hz,Ar-H),7.83(s,1H,Ar-H),7.52(d,1H,J=8.3Hz,Ar-H),7.44(d,1H,J=8.3Hz,Ar-H),7.10(t,1H,J=7.4Hz,Ar-H),7.05(t,1H,J=7.7Hz,Ar-H),6.98(d,1H,J=7.7Hz,Ar-H),6.78(t,1H,J=7.6Hz,Ar-H),6.77(d,1H,J=7.6Hz,Ar-H),6.70(t,1H,J=7.1Hz,Ar-H),4.25(q,2H,J=7.3Hz,-CH2 -CH3),1.30(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ167.1,167.0,136.8,136.2,133.1,131.3,128.7,127.9,126.1,125.2,122.7×2,122.2,121.9,120.7,120.5,112.8,111.0,105.5,104.8,41.4,15.7.ESI-MS m/z 357.1[M+H]+.
vi)N-(N,N-二甲氨基乙基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(24)的制备
以30mL甲苯溶解化合物24d(55mg,0.154mmol)、N,N-二甲基乙二胺(84.4μL,0.772mmol)和催化量Et3N,甲苯溶解后,在氮气保护下,110℃冷凝水回流17h,蒸干溶剂,加压柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得红色固体(24)52mg,收率79%。1H NMR(600MHz,DMSO-d6)δ11.73(s,1H,indole-NH),7.80(d,1H,J=2.7Hz,Ar-H),7.77(s,1H,Ar-H),7.47(d,1H,J=8.3Hz,Ar-H),7.39(d,1H,J=8.1Hz,Ar-H),7.05(t,1H,J=7.7Hz,Ar-H),6.99(t,1H,J=7.6Hz,Ar-H),6.89(d,1H,J=7.9Hz,),6.74(d,1H,J=8.1Hz,Ar-H),6.70(d,1H,J=7.4Hz,Ar-H),6.62(d,1H,J=7.8Hz,Ar-H),4.24(q,2H,J=7.2Hz,-CH2 -CH3),3.66(t,2H,J=6.4Hz,N-CH2 -CH2N(CH3)2),2.49(t,J=6.4Hz,2H,NCH2-CH2 -N(CH3)2),2.18(s,6H,-N(CH3)2),1.31(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ172.1×2,136.6,136.0,132.1,130.0,127.4,126.8,126.5,125.6,122.3,122.2,121.9,121.6,120.2,120.0,112.4,110.7,106.1,105.5,57.3,45.7×2,41.3,36.3,15.8.HR-ESIMS m/z427.2140[M+H]+(calcd.for C26H27N4O2,427.2134).
化合物25的制备
按照化合物16的制备方法,以化合物24(50mg,0.117mmol)为原料得其盐酸盐:N-(N,N-二甲基氨乙基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺盐酸盐(25)(48mg,收率90%)。1H NMR(500MHz,DMSO-d6)δ11.94(s,1H,indole-NH),10.61(brs,1H,(CH3)2NH +),7.77(d,1H,J=2.4Hz,Ar-H),7.69(s,1H,Ar-H),7.47(d,1H,J=8.2Hz,Ar-H),7.40(d,1H,J=8.1Hz,Ar-H),7.05(t,1H,J=7.6Hz,Ar-H),6.99(d,1H,J=7.8Hz,Ar-H),6.98(t,1H,J=7.8Hz,Ar-H),6.76(d,1H,J=8.0Hz,Ar-H),6.72(t,1H,J=7.5Hz,Ar-H),6.62(t,1H,J=7.5Hz,Ar-H),4.23(q,2H,J=7.0Hz,-CH2 -CH3),3.93(t,2H,J=5.7Hz,N-CH 2-CH2(CH3)2NH+),3.36(t,2H,J=6.0Hz,NCH2-CH2 -(CH3)2NH+),2.84(s,6H,(CH3)2 NH+),1.28(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.9×2,136.5,135.9,131.8,129.9,128.0,127.1,126.4,125.3,122.2,122.1×2,121.7,120.0,119.8,112.4,110.9,105.9,105.5,54.8,42.6×2,41.1,33.9,15.7.HR-ESIMS m/z 427.2141[M–Cl]+(calcd.for C26H26N4O2,426.2056).
化合物26的制备
i)1-苄基-6-溴吲哚(26a)的制备
按照化合物24a的合成方法,以6-溴吲哚(980mg,5mmol)、NaH(300mg,7.5mmol,质量分数60%,分散在石蜡中)和溴化苄(1283mg,7.5mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=60:1(v/v)洗脱白色粉末状固体(26a)1.14g,收率80%。1H NMR(600MHz,CDCl3)δ7.54(d,1H,J=8.8Hz,Ar-H),7.47(s,1H,Ar-H),7.35–7.31(m,3H,Ar-H),7.25(dd,1H,J=8.8Hz,1.9Hz,Ar-H),7.10–7.12(m,3H,Ar-H),6.56(d,1H,J=3.8Hz,Ar-H),5.27(s,2H,Ph-CH2 -).13C NMR(150MHz,CDCl3)δ137.3,137.1,129.0×2,127.9,127.7,126.8×2,123.0,122.3,115.5,112.8,102.1,50.2.ESI-MS m/z286.0/288.0[M+H]+.
ii)2-(1-乙基-3-吲哚)-3-(1-苄基-6-溴-3-吲哚)马来酸酐(26b)的制备
按照化合物24b的制备方法,以化合物24a(1100mg,3.86mmol)、(COCl)2(500μL,5.79mmol)、化合物1c(783mg,3.86mmol)和Et3N(1070μL,7.72mmol)为原料制备,甲醇重结晶得红色粉末(26b)652mg,收率32.2%。1H NMR(600MHz,DMSO-d6)δ8.02(s,1H,Ar-H),7.99(s,1H,Ar-H),7.73(s,1H,Ar-H),7.54(d,1H,J=8.2Hz,Ar-H),7.35(t,2H,J=7.7Hz,Ar-H),7.29(t,1H,J=6.6Hz,Ar-H),7.19(d,2H,J=7.1Hz,Ar-H),7.12(t,1H,J=7.7Hz,Ar-H),6.90(d,1H,J=8.2Hz,Ar-H),6.88(d,1H,J=7.7Hz,Ar-H),6.76(d,1H,J=7.7Hz,Ar-H),6.71(t,1H,J=7.7Hz,Ar-H),5.53(s,2H,Ph-CH2-),4.32(q,2H,J=7.2Hz,-CH2 -CH3),1.37(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.9,166.8,137.6,137.3,136.3,134.4,133.6,130.0,129.2×2,128.2,127.5×2,126.8,125.7,125.4,123.6,123.5,122.9,122.0,120.8,115.8,114.3,111.2,105.5,104.5,50.0,41.6,15.8.ESI-MSm/z 525.1/527.1[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-6-溴-3-吲哚)马来酰亚胺(26c)制备
按照化合物24c的制备方法,以化合物26b(600mg,1.14mmol)、HMDS(12mL,57.3mmol)和MeOH(1.2mL,28.7mmol)为原料制备,经硅胶柱色谱分离、二氯甲烷洗脱得红色粉末状固体(26c)218mg,收率95%。1H NMR(600MHz,DMSO-d6)δ10.99(s,1H,imide-NH),7.90(s,1H,Ar-H),7.89(s,1H,Ar-H),7.65(s,1H,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.34(t,2H,J=7.7Hz,Ar-H),7.28(t,1H,J=7.1Hz,Ar-H),7.17(d,2H,J=7.1Hz,Ar-H),7.05(t,1H,J=7.7Hz,Ar-H),6.82(d,1H,J=8.8Hz,Ar-H),6.79(d,1H,J=7.1Hz,Ar-H),6.72(d,1H,J=8.3Hz,Ar-H),6.63(t,1H,J=7.1Hz,Ar-H),5.51(s,2H,Ph-CH2-),4.29(q,2H,J=7.1Hz,-CH2 -CH3),1.37(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.3,173.1,137.9,137.2,136.1,133.4,132.3,129.3,129.2×2,128.1,127.4,127.4,126.5,126.2,125.9,123.3,123.0,122.3,121.7,120.2,115.3,113.9,110.8,106.3,105.1,49.8,41.3,15.8.ESI-MS m/z 524.1/526.1[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(26d)的制备
按照化合物24d的制备方法,以化合物26c(538mg,1.03mmol)、DMSO(1.7mL)、1M的t-BuOK/THF溶液(16.8mL,16.8mmol)和O2为原料制备。硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末(26d)366mg,收率82%。1H NMR(600MHz,DMSO-d6)δ7.81(s,1H,Ar-H),7.76(d,1H,J=2.8Hz,Ar-H),7.57(s,1H,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.04(dt,1H,J=8.2Hz,1.1Hz,Ar-H),6.78(d,1H,J=8.0Hz,Ar-H),6.75(dd,1H,J=8.6Hz,1.8Hz,Ar-H),6.71(d,1H,J=8.8Hz,Ar-H),6.69(dt,1H,J=7.3Hz,0.9Hz,Ar-H),4.26(q,2H,J=7.1Hz,-CH2 -CH3),1.34(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.4×2,137.4,136.0,132.2,130.6,128.6,127.4,126.4,125.0,123.1,122.6,122.3,121.7,120.2,115.0,114.9,110.7,106.3,105.3,41.3,15.8.ESI-MS m/z434.0/436.0[M+H]+,HR-ESIMS m/z 434.0506[M+H]+(calcd.for C22H17N3O2Br,434.0504).
v)2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酸酐(26e)的制备
按照化合物24e的制备方法,由化合物26d(80mg,0.185mmol)为原料制备,得橙红色固体(26e)60mg,收率75%。1H NMR(600MHz,DMSO-d6)δ11.99(s,1H,indole-NH),7.88(d,1H,J=2.7Hz,Ar-H),7.84(s,1H,Ar-H),7.62(s,1H,Ar-H),7.49(d,1H,J=8.0Hz,Ar-H),7.08(t,1H,J=7.6Hz,Ar-H),6.80(t,2H,J=7.1Hz,Ar-H),6.73(t,2H,J=7.5Hz,Ar-H),4.26(q,2H,J=8.2Hz,-CH2 -CH3),1.32(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.8×2,137.4,136.1,133.2,131.7,128.9,127.5,125.8,124.4,123.2,123.0,122.7,121.9,120.7,115.7,115.2,111.0,105.5,104.5,41.4,15.8.ESI-MS m/z 435.0/437.0[M+H]+.
vi)N-(N,N-二甲氨基乙基)-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(26)的制备
按照化合物24的制备方法,以化合物26e(50mg,0.115mmol)、N,N-二甲基乙二胺(81μL,0.575mmol)和催化量Et3N为原料制备,得红色固体(26)46.4mg,收率80%。1H NMR(500MHz,DMSO-d6)δ11.79(s,1H,indole-NH),7.84(s,1H,Ar-H),7.77(s,1H,Ar-H),7.56(d,1H,J=1.4Hz,Ar-H),7.47(d,1H,J=8.2Hz,Ar-H),7.03(t,1H,J=7.6Hz,Ar-H),6.74(d,2H,J=8.5Hz,Ar-H),6.69(d,1H,J=6.5Hz,Ar-H),6.68(t,1H,J=62Hz,Ar-H),4.26(q,2H,J=7.2Hz,-CH2 -CH3),3.64(t,2H,J=6.4Hz,-NCH2 CH2-N(CH3)2),2.46(t,2H,J=6.5Hz,-NCH2CH2 -N(CH3)2),2.16(s,6H,-N(CH3)2),1.33(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.8×2,137.3,136.0,132.2,130.5,127.6,126.3,126.2,124.8,122.9,122.6,122.3,121.6,120.1,114.9,114.8,110.7,106.2,105.1,57.2,45.6,41.2,36.2,15.7.HR-ESIMS m/z 505.1250[M+H]+(calcd.for C26H26N4O2Br,505.1239).
化合物27的制备
按照化合物16的制备方法,以化合物26(100mg,0.198mmol)为原料得其盐酸盐:N-(N,N-二甲基氨乙基)-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺盐酸盐(27)(91mg,收率85%)。1H NMR(500MHz,DMSO-d6)δ12.08(s,1H,indole-NH),10.63(brs,1H,-(CH3)2NH +),7.79(s,1H,Ar-H),7.74(s,1H,Ar-H),7.60(s,1H,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.05(t,1H,J=7.5Hz,Ar-H),6.84(d,1H,J=8.0Hz,Ar-H),6.76(d,1H,J=7.2Hz,Ar-H),6.74(s,1H,Ar-H),6.70(d,1H,J=7.4Hz,Ar-H),4.26(q,2H,J=6.9Hz,-CH2 -CH3),3.92(t,2H,J=4.9Hz,N-CH 2-CH2N(CH3)2H+),3.04(t,2H,J=4.9Hz,NCH2-CH2 -N(CH3)2H+),2.83(s,6H,-(CH3)2 NH+),1.32(t,3,J=6.9Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.8,171.6,137.4,136.0,132.1,130.5,128.0,126.9,126.1,124.7,123.1,122.5,122.3,121.8,120.1,115.0,114.8,110.7,106.2,105.2,54.8,45.6,42.6×2,41.2,33.5,15.7.HR-ESIMS m/z 505.1246[M–Cl]+(calcd.for C26H27N4O2Br,505.1250).
化合物28的制备
按照化合物14的制备方法,以化合物26e(49mg,0.114mmol)和乙二胺为原料制得红色固体N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(28)49mg,收率90%。1H NMR(500MHz,DMSO-d6)δ7.81(s,1H,Ar-H),7.76(s,1H,Ar-H),7.56(s,1H,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.03(t,1H,J=7.6Hz,Ar-H),6.77(d,1H,J=8.0Hz,Ar-H),6.74(dd,1H,J=8.6Hz,1.4Hz,Ar-H),6.70(d,1H,J=8.6Hz,Ar-H),6.67(t,1H,J=7.5Hz,Ar-H),4.25(q,2H,J=7.2Hz,-CH2 -CH3),3.56(t,2H,J=6.5Hz,N-CH2 -CH2NH2),2.77(t,2H,J=6.5Hz,NCH2-CH2 -NH2),1.32(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.1×2,137.3,135.9,132.0,130.4,127.7,126.4,126.2,124.8,123.0,122.5,122.2,121.6,120.1,114.9,114.8,110.7,106.3,105.2,41.3,41.2,40.6,15.7.HR-ESIMS m/z 477.0934[M+H]+(calcd.for C24H22N4O2Br,477.0926).
化合物29的制备
按照化合物16的制备方法,以化合物28(200mg,0.42mmol)为原料得其盐酸盐:N-(2-氨乙基)-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺盐酸盐(29)172mg,收率80%。1H NMR(500MHz,DMSO-d6)δ12.03(s,1H,indole-NH),8.20(brs,3H,-NH3 +),7.80(s,1H,Ar-H),7.75(s,1H,Ar-H),7.60(s,1H,Ar-H),7.48(d,1H,J=8.1Hz,Ar-H),7.05(t,1H,J=7.4Hz,Ar-H),6.83(d,1H,J=7.9Hz,Ar-H),6.76(d,1H,J=8.5Hz,Ar-H),6.73(d,1H,J=8.8Hz,Ar-H),6.72(t,1H,J=7.8Hz,Ar-H),4.26(q,2H,J=6.9Hz,-CH2 -CH3),3.83(t,2H,J=6.4Hz,N-CH2 -CH2NH3 +),3.08(t,2H,J=6.5Hz,NCH2-CH2 -NH3 +),1.32(t,3H,J=6.9Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.9,171.8,137.4,136.9,132.0,130.5,127.9,126.8,126.1,124.7,123.1,122.5,122.3,121.8,120.1,115.0,114.8,110.7,106.2,105.2,41.2,38.1,36.1,15.7.HR-ESIMS m/z 477.0932[M–Cl]+(calcd.for C24H22N4O2Br,477.0934).
化合物30、31、32的制备
0℃下,将NaH(13.5mg,0.563mmol,质量分数60%,分散于石蜡中)以DMF悬浮,滴加DMF溶解的化合物24d(40mg,0.113mmol),以DMF悬浮NaH(338mg,14.1mmol,质量分数60%,分散于石蜡中),滴加氯代乙醇(38μL,0.563mmol),低温反应20min,以导管将其导入至化合物24d的悬浮液中,反应20min后,升至室温,冷凝水回流4.5h。降温至–5℃,滴加MeOH,加入适量饱和氯化铵溶液,乙酸乙酯萃取,合并有有机层,并用无水硫酸钠干燥,真空蒸干溶剂,加压柱色谱分离、二氯甲烷:甲醇=100:1(v/v)洗脱得红色固体N-(2-羟乙基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(30)、2-(1-乙基-3-吲哚)-3-(1-(2-羟乙基)-3-吲哚)马来酰亚胺(31)和N-(2-羟乙基)-2-(1-乙基-3-吲哚)-3-(1-(2-羟乙基)-3-吲哚)马来酰亚胺(32)分别为23mg、5mg和4mg,收率分别为51%、11%和8%。
化合物30:1H NMR(600MHz,DMSO-d6)δ11.67(s,1H,indole-NH),7.78(d,1H,J=2.1Hz,Ar-H),7.73(s,1H,Ar-H),7.45(d,1H,J=8.2Hz,Ar-H),7.37(d,1H,J=8.1Hz,Ar-H),7.03(t,1H,J=7.6Hz,Ar-H),6.97(t,1H,J=7.6Hz,Ar-H),6.91(d,1H,J=8.0Hz,Ar-H),6.73(d,1H,J=8.0Hz,Ar-H),6.68(t,1H,J=7.5Hz,Ar-H),6.61(t,1H,J=7.6Hz,Ar-H),4.89(t,1H,J=5.5Hz,imide-NCH2CH2OH),4.22(q,2H,J=7.2Hz,-CH2 -CH3),3.62-3.59(m,4H,imide-N(CH2)2-),1.29(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ172.1×2,136.5,135.9,131.8,129.7,127.5,126.8,126.4,125.5,122.1×2,121.8,121.5,120.0,119.8,112.2,110.5,106.0,105.5,58.7,41.1,40.9,15.6.HR-ESIMS m/z400.1666[M+H]+(calcd.for C24H22N3O3,400.1661).
化合物31:1H NMR(600MHz,DMSO-d6)δ10.92(s,1H,imide-NH),7.82(s,1H,Ar-H),7.70(s,1H,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.46(d,1H,J=8.2Hz,Ar-H),7.05(t,1H,J=7.6Hz,Ar-H),7.02(t,1H,J=7.1Hz,Ar-H),6.92(d,1H,J=8.0Hz,Ar-H),6.72(d,1H,J=7.3Hz,Ar-H),6.71(t,1H,J=6.9Hz,Ar-H),6.64(t,1H,J=7.7Hz,Ar-H),4.95(t,1H,J=5.2Hz,indole-NCH2CH2OH),4.26(t,2H,J=5.6Hz,indole-NCH2 CH2-),4.23(q,2H,J=7.2Hz,-CH2 -CH3)3.71-3.69(m,2H,indole-NCH2CH2 -),1.30(t,3H,J=7.2Hz,-CH2-CH3 ).13CNMR(150MHz,DMSO-d6)δ173.5×2,136.7,136.0,133.4,131.9,128.0,127.5,126.9,126.1,122.2,122.1,122.0,121.9,120.1,120.0,111.0,110.6,105.5,105.2,60.6,49.1,41.2,15.8.HR-ESIMS m/z 400.1668[M+H]+(calcd.for C24H22N3O3).
化合物32:1H NMR(500MHz,DMSO-d6)δ7.84(s,1H,Ar-H),7.72(s,1H,Ar-H),7.48(d,1H,J=7.8Hz,Ar-H),7.46(d,1H,J=7.8Hz,Ar-H),7.05(t,1H,J=7.2Hz,Ar-H),7.02(t,1H,J=7.4Hz,Ar-H),6.93(d,1H,J=8.0Hz,Ar-H),6.73(d,1H,J=7.8Hz,Ar-H),6.71(t,1H,J=7.5Hz,Ar-H),6.65(t,1H,J=7.3Hz,Ar-H),4.93(t,1H,J=4.7Hz,indole-NCH2CH2OH),4.88(t,1H,J=4.8Hz,imide-NCH2CH2OH),4.26(t,2H,J=5.3Hz,indole-NCH2 CH2-),4.22(q,2H,J=7.1Hz,-CH2 -CH3),3.70(t,2H,J=5.3Hz,indole-NCH2CH2 -),3.65-3.57(m,4H,imide-N(CH2)2-),1.29(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.0×2,136.6,135.9,133.3,131.8,127.1,126.7,126.4,125.9,122.1,122.0×2,121.8,120.0,119.9,110.9,110.5,105.9,105.2,60.5,58.6,49.0,41.1,40.9,15.6.HR-ESIMS m/z 444.1934(calcd.for C26H26N3O4,444.1923[M+H]+).
化合物33的制备
按照化合物24的制备方法,以4-甲氧基苄胺(185μL,1.4mmol)和化合物24e(50mg,0.14mmol)为原料得N-(4-甲氧基苄基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(33)48mg,收率72%。1H NMR(600MHz,DMSO-d6)δ11.75(s,1H,indole-NH),7.82(d,1H,J=2.7Hz,Ar-H),7.77(s,1H,Ar-H),7.45(d,1H,J=8.3Hz,Ar-H),7.39(d,1H,J=8.1Hz,Ar-H),7.29(d,2H,J=8.7Hz,Ar-H),7.04(t,1H,J=7.6Hz,Ar-H),6.98(t,1H,J=7.6Hz,Ar-H),6.90(d,2H,J=6.7Hz,Ar-H),6.89(d,1H,J=2.1Hz,Ar-H),6.74(d,1H,J=8.1Hz,Ar-H),6.69(t,1H,J=7.5Hz,Ar-H),6.62(t,1H,J=7.5Hz,Ar-H),4.68(s,2H,-CH2 -),4.24(q,2H,J=7.2Hz,-CH2 -CH3),3.70(s,3H,-OCH3),1.29(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ172.0,171.9,159.1,136.6,136.0,132.2,130.1,129.9,129.6×2,127.5,127.3,126.9,126.5,125.6,122.3×2,121.9,121.6,120.2,112.0,114.5,112.4,110.7,106.1,105.5,55.6,41.2,41.1,15.7.ESI-MS m/z 498.2[M+Na]+.
化合物34的制备
按照化合物2的制备方法,以化合物33(60mg,0.126mmol)和NaHCO3(42mg,0.5mmol)和HCHO(3mL,质量分数37%)溶液为原料制备,凝胶柱色谱分离、甲醇洗脱得深红色固体N-(4-甲氧基苄基)-2-(1-乙基-3-吲哚)-3-(1-羟甲基-3-吲哚)马来酰亚胺(34)51mg,收率78%。1H NMR(600MHz,DMSO-d6)δ7.99(s,1H,Ar-H),7.74(s,1H,Ar-H),7.55(d,1H,J=8.3Hz,Ar-H),7.47(d,1H,J=8.2Hz,Ar-H),7.39(t,1H,J=8.1Hz,Ar-H),7.29(d,2H,J=8.8Hz,Ar-H),7.05(m,2H,Ar-H),6.99(d,1H,J=8.1Hz,Ar-H),6.91(d,2H,J=8.8Hz,Ar-H),6.72(d,1H,J=7.2Hz,Ar-H),6.69(t,1H,J=8.1Hz,-CH2OH),6.63(d,1H,J=7.0Hz,Ar-H),5.60(d,2H,J=7.2Hz,-CH2 OH),4.70(s,2H,-CH2-),4.23(q,2H,J=7.2Hz,-CH2 -CH3),3.71(s,3H,-OCH3),1.30(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.9,171.8,159.2,136.1,136.0,132.6×2,132.2,129.8,129.6,127.4,126.9,126.8,126.4,122.4×2,121.9,121.8,120.5,120.3,114.6,111.4,110.7,105.9,105.5,100.0,69.7,60.3,41.7,41.3,15.7.ESI-MS m/z 528.1[M+Na]+.
化合物35的制备
按照化合物24的制备方法,以4-羟基苄胺(86mg,0.7mmol)和化合物24e(50mg,0.14mmol)为原料,制得N-(4-羟基苄基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(35)35mg,收率60%。1H NMR(600MHz,DMSO-d6)δ11.72(s,1H,indole-NH),9.26(s,1H,Ar-OH),7.80(d,1H,J=2.3Hz,Ar-H),7.77(s,1H,Ar-H),7.45(d,1H,J=8.3Hz,Ar-H),7.38(d,1H,J=8.1Hz,Ar-H),7.17(d,1H,J=8.5Hz,Ar-H),7.11(d,2H,J=8.3Hz,Ar-H),7.03(t,1H,J=7.4Hz,Ar-H),6.97(d,1H,J=7.5Hz,Ar-H),6.89(d,1H,J=8.0Hz,Ar-H),6.71(d,2H,J=8.4Hz,Ar-H),6.68(d,1H,J=7.4Hz,Ar-H),6.61(t,1H,J=7.4Hz,Ar-H),4.64(s,2H,-NCH2Ar),4.22(q,2H,J=7.2Hz,-CH2 -CH3),1.30(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ172.0×2,156.7,136.6,136.0,134.1,132.1,132.0,130.1,129.9,129.7,129.2,128.1,127.9,127.5,126.9,126.5,125.6,115.8×2,115.5×2,112.4,110.7,106.1,105.5,56.6,41.2,15.7.ESI-MS m/z 462.3[M+H]+.
化合物36的制备
按照化合物2的制备方法,以化合物35(59mg,0.128mmol)和NaHCO3(53.7mg,0.64mmol)为原料,制得深红色固体N-(4-羟基苄基)-2-(1-乙基-3-吲哚)-3-(1-羟甲基-3-吲哚)马来酰亚胺(36)50mg,收率80%。1H NMR(600MHz,DMSO-d6)δ9.44(s,1H,Ar-OH),8.00(s,1H,Ar-H),7.75(s,1H,Ar-H),7.56(d,1H,J=8.3Hz,Ar-H),7.47(d,1H,J=8.3Hz,Ar-H),7.19(d,2H,J=8.5Hz,Ar-H),7.05(m,2H,Ar-H),7.00(d,1H,J=8.1Hz,Ar-H),6.74(d,2H,J=8.6Hz,Ar-H),6.72(d,1H,J=7.4Hz,Ar-H),6.69(t,1H,J=7.4Hz,-CH2OH),6.63(d,1H,J=3.3Hz,Ar-H),5.61(d,2H,J=7.4Hz,-CH2OH),4.65(s,2H,-NCH2Ar),4.03(q,2H,J=7.1Hz,-CH2 -CH3),1.17(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.9×2,157.3,136.1,136.0,132.7,132.6,132.2,129.7,128.1,127.3,126.8×2,126.4,122.4×2,121.9,121.8,120.5,120.3,115.8,111.4,110.7,105.9,105.5,100.0,69.7,60.3,41.3,15.7.ESI-MS m/z 514.1[M+Na]+.
化合物37的制备
按照化合物24的制备方法,以4-(2-氨乙基)吗啉(771μL,0.59mmol)和化合物24e(50mg,0.14mmol)为原料,制得N-(2-(4-吗啉)乙基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(37)41mg,收率61%。1H NMR(500MHz,DMSO-d6)δ11.68(s,1H,indole-NH),7.78(s,1H,Ar-H),7.73(s,1H,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.37(d,1H,J=8.1Hz,Ar-H),7.04(t,1H,J=7.6Hz,Ar-H),6.98(t,1H,J=7.5Hz,Ar-H),6.90(d,1H,J=8.0Hz,Ar-H),6.71(d,1H,J=8.3Hz,Ar-H),6.70(t,1H,J=7.5Hz,Ar-H),6.62(t,1H,J=7.5Hz,Ar-H),4.23(q,2H,J=7.2Hz,-CH2 -CH3),3.68(t,2H,J=6.4Hz,imide-NCH2 CH2-),3.53(t,4H,J=4.4Hz,morpholine-N(CH2-CH 2)2O),2.53(t,2H,J=6.4Hz,imide-NCH2CH2 -),2.44(t,4H,J=4.4Hz,morpholine-N(CH 2-CH2)2O),1.30(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.0×2,136.5,135.9,131.9,129.8,127.4,126.8,126.4,125.4,122.2,122.1,121.7,121.4,120.0,119.8,112.3,110.5,106.0,105.3,66.7×2,56.3,53.6×2,41.1,35.2,15.6.HR-ESIMS m/z 469.2247[M+H]+(calcd.for C28H29N4O3,469.2240[M+H]+).
化合物38的制备
按照化合物2的制备方法,以化合物37(33mg,0.071mmol)和NaHCO3(30mg,0.35mmol)为原料,制得深红色固体N-(2-(4-吗啉)乙基)-2-(1-乙基-3-吲哚)-3-(1-羟甲基-3-吲哚)马来酰亚胺(38)29mg,收率82%。1H NMR(500MHz,DMSO-d6)δ7.96(s,1H,Ar-H),7.70(s,1H,Ar-H),7.55(d,1H,J=8.2Hz,Ar-H),7.47(d,1H,J=8.2Hz,Ar-H),7.05(t,1H,J=8.1Hz,Ar-H),7.02(t,1H,J=8.4Hz,Ar-H),6.73(t,1H,J=7.5Hz,Ar-H),6.67(t,1H,J=7.3Hz,Ar-H),6.62(d,1H,J=8.5Hz,Ar-H),6.60(d,1H,J=8.0Hz,Ar-H),5.59(d,2H,J=7.3Hz,indole-CH2 -OH),4.22(q,2H,J=7.2Hz,-CH2 -CH3),3.72(t,2H,J=6.4Hz,imide-NCH2 CH2-),3.56(t,4H,J=4.5Hz,morpholine-N(CH2-CH 2)2O),2.57(t,2H,J=6.4Hz,imide-NCH2CH2 -),2.47(t,4H,J=4.5Hz,morpholine-N(CH 2-CH2)2O),1.29(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.9×2,136.0,135.9,132.4,131.9,127.3,126.9,126.6,126.2,122.3,122.2,121.8,121.6,120.3,120.1,111.9,110.6,105.7,105.4,69.5,66.6×2,56.3,53.5×2,41.1,35.3,15.6.HR-ESIMS m/z 499.2352[M+H]+(calcd.forC29H31N4O4,499.2345).
化合物39的制备
按照化合物24的制备方法,以2-(2-氨乙基)吡啶(50μL,0.421mmol)和化合物24e(30mg,0.084mmol)为原料,制得N-(2-(2-吡啶)乙基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(39)25mg,收率64%。1H NMR(600MHz,DMSO-d6)δ11.71(s,1H,indole-NH),8.48(d,1H,J=4.7Hz,Ar-H),7.75(d,1H,J=2.7Hz,Ar-H),7.73–7.70(m,2H,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.38(d,1H,J=8.1Hz,Ar-H),7.31(d,1H,J=7.7Hz,Ar-H),7.23(dd,1H,J=7.4,4.9Hz,Ar-H),7.05(t,1H,J=7.6Hz,Ar-H),6.99(t,1H,J=7.5Hz,Ar-H),6.88(d,1H,J=8.1Hz,Ar-H),6.71(t,2H,J=8.0Hz,Ar-H),6.64(d,1H,J=7.7Hz,Ar-H),4.24(q,2H,J=7.2Hz,-CH2 -CH3),3.93(t,2H,J=7.1Hz,imide-NCH2 CH2-),3.09(t,2H,J=7.1Hz,imide-NCH2CH2 -),1.30(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ172.0,171.9,158.9,149.7,137.2,136.6,136.0,132.0,129.9,127.6,126.9,126.5,125.6,123.8,122.3,122.2,121.9,121.6,120.1,119.9,112.4,110.7,106.0,105.5,100.0,41.2,38.2,36.7,15.8.HR-ESIMS m/z 461.1981[M+H]+(calcd.for C29H25N4O2,461.1978).
化合物40的制备
按照化合物16的制备方法,以化合物39(200mg,0.435mmol)为原料制得其盐酸盐:N-(2-(2-吡啶)乙基)-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺盐酸盐(40)173mg,收率80%。1H NMR(500MHz,DMSO-d6)δ11.88(s,1H,indole-NH),10.60(brs,1H,-pryidine-H+),8.82(s,1H,Ar-H),8.45(t,1H,J=7.2Hz,Ar-H),7.99(d,1H,J=6.4Hz,Ar-H),7.87(t,1H,J=7.0Hz,Ar-H),7.68(d,1H,J=7.1Hz,Ar-H),7.61(d,1H,J=7.6Hz,Ar-H),7.45(d,1H,J=7.7Hz,Ar-H),7.37(d,1H,J=7.4Hz,Ar-H),7.04(t,1H,J=6.8Hz,Ar-H),6.97(t,1H,J=7.1Hz,Ar-H),6.87(d,1H,J=7.5Hz,Ar-H),6.70(t,1H,J=6.6Hz,Ar-H),6.65(d,1H,J=7.1Hz,Ar-H),6.61(d,1H,J=6.6Hz,Ar-H),4.20(q,2H,J=7.1Hz,-CH2 -CH3),4.02(t,2H,J=7.1Hz,imide-NCH2 CH2-),3.39(t,2H,J=7.1Hz,imide-NCH2CH2 -),1.26(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.7×2,155.0,145.7,142.4,136.5,135.9,131.8,129.9,127.9,127.6,126.7,126.3,125.4,125.9,122.2,122.1,121.9,121.6,120.0,119.8,112.3,110.6,105.8,105.3,45.7,41.1,37.6,15.6.HR-ESIMS m/z 461.1989[M-Cl]+(calcd.for C29H25N4O2,461.1978).
化合物41的制备
以10mL THF溶解化合物24e(52mg,0.146mmol),滴加水合肼(73μL,1.46mmol)。反应液加热到45℃反应15min,真空蒸干溶剂,加水与乙酸乙酯萃取,有机层蒸干,硅胶柱色谱分离、二氯甲烷:甲醇=100:1(v/v)洗脱得红色固体N-氨基-2-(1-乙基-3吲哚)-3-(3-吲哚)马来酰亚胺(41)45mg,收率83.3%。1H NMR(600MHz,DMSO-d6)δ11.71(d,1H,J=2.2Hz,indole-NH),7.81(d,1H,J=2.2Hz,Ar-H),7.73(s,1H,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.40(d,1H,J=8.3Hz,Ar-H),7.05(t,1H,J=7.1Hz,Ar-H),6.99(t,1H,J=7.1Hz,Ar-H),6.93(d,1H,J=7.7Hz,Ar-H),6.74(d,1H,J=8.2Hz,Ar-H),6.71(t,1H,J=7.7Hz,Ar-H),6.63(t,1H,J=7.7Hz,Ar-H),4.85(s,2H,-NH2),4.22(q,2H,J=7.7Hz,-CH2 -CH3),1.30(t,3H,J=7.7Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.5,171.4,136.6,136.0,131.9,129.9,126.6,126.1,125.6,125.4,122.3,122.2,121.9,121.5,120.1,119.9,112.4,110.7,106.2,105.6,41.2,15.8.HR-ESIMS m/z 371.1511[M+H]+(calcd.for C22H19N4O2,371.1508).
化合物42的制备
在10mL单口瓶中,以HCHO(4mL,质量分数37%)悬浮化合物24d(22mg,0.05mmol),室温反应过夜。TLC检测至反应完毕,倒入冰水中(30mL),乙酸乙酯萃取(2×50mL),饱和食盐水(2×50mL),合并有机相,无水硫酸钠干燥,真空旋蒸除去溶剂,以硅胶柱色谱分离、二氯甲烷:甲醇=80:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(3-吲哚)马来酰亚胺(42)12mg,收率60%。1H NMR(500MHz,DMSO-d6)δ11.71(s,1H,indole-NH),7.81(d,1H,J=2.7Hz,Ar-H),7.76(s,1H,Ar-H),7.48(d,1H,J=8.2Hz,Ar-H),7.39(d,1H,J=8.1Hz,Ar-H),7.06(t,1H,J=7.0Hz,Ar-H),6.99(t,1H,J=7.4Hz,Ar-H),6.92(d,1H,J=8.0Hz,Ar-H),6.73(dd,2H,J=7.7Hz,4.6Hz,Ar-H),6.64(d,1H,J=7.4Hz,Ar-H),6.30(t,1H,J=7.0Hz,-CH2-OH),4.97(d,2H,J=7.0Hz,-CH2 -OH),4.25(q,2H,J=7.2Hz,-CH2 -CH3),1.32(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.7×2,136.6,136.1,132.1,130.1,128.0,127.3,126.6,125.6,122.3×2,121.9,121.5,120.2,120.0,112.4,110.7,106.0,105.4,60.8,41.6,15.7.HR-ESIMS m/z 386.1490[M+H]+(calcd.forC23H20N3O3,386.1499[M+H]+).
化合物43的制备
i)1-苄基-6-氟吲哚(43a)的制备
按照化合物24a的合成方法,以6-氟吲哚(675mg,5mmol)、NaH(300mg,7.5mmol,质量分数60%,分散于石蜡中)和溴化苄(1283mg,7.5mmol)为原料得白色结晶粉末(43a)1.01g,收率90%。1H NMR(600MHz,CDCl3)δ7.53(dd,1H,J=8.7Hz,5.5Hz,Ar-H),7.29-7.24(m,3H,Ar-H),7.06-7.09(m,3H,Ar-H),6.92(dd,1H,J=9.6Hz,2.3Hz,Ar-H),6.86(dt,1H,J=9.6Hz,2.3Hz,Ar-H),6.51(d,1H,J=3.2Hz,Ar-H),5.21(s,2H,-CH2-Ph).13C NMR(150MHz,CDCl3)δ159.9(d,1JCF=240Hz),137.2,136.5(d,3JCF=12Hz),129.0,128.9×2,127.9,126.9×2,125.3,121.7(d,3JCF=10Hz),108.5(d,2JCF=25Hz),102.0,96.3(d,2JCF=26Hz),50.4.ESI-MS m/z 225.1[M+H]+.
ii)2-(1-乙基-3-吲哚)-3-(1-苄基-6-氟-3-吲哚)马来酸酐(43b)的制备
按照化合物24b的合成方法,以化合物43a(600mg,2.67mmol)、(COCl)2(345μL,4.00mmol)、化合物1a(541mg,2.67mmol)和Et3N(738μL,5.33mmol)为原料,制得红色固体(43b)400mg,收率32%。1H NMR(600MHz,DMSO-d6)δ8.00(s,1H,Ar-H),7.99(s,1H,Ar-H),7.53(d,1H,J=8.3Hz,Ar-H),7.36(dd,1H,J=8.7Hz,2.3Hz,Ar-H),7.34(t,2H,J=7.3Hz,Ar-H),7.28(t,1H,J=7.3Hz,Ar-H),7.22(d,2H,J=7.3Hz,Ar-H),7.11(t,1H,J=6.9Hz,Ar-H),6.89(dd,1H,J=8.7Hz,5.5Hz,Ar-H),6.78(d,1H,J=7.8Hz,Ar-H),6.69(t,1H,J=7.8Hz,Ar-H),6.63(dt,1H,J=9.2Hz,2.3Hz,Ar-H),5.49(s,2H,-CH2-Ph),4.32(q,2H,J=7.3Hz,-CH2 -CH3),1.36(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.9,166.8,155.9(d,1JCF=235Hz),137.6,136.6(d,3JCF=12Hz),136.3,134.3,133.5,129.6,129.2×2,128.2,127.6×2,127.1,125.8,123.1(d,3JCF=13Hz),123.0,122.8,122.1,120.7,111.2,109.2(d,2JCF=23Hz),105.5,104.6,98.0(d,2JCF=26Hz),50.0,41.5,15.8.ESI-MS m/z 465.2[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-6-氟-3-吲哚)马来酰亚胺(43c)的制备
按照化合物24c的制备方法,以化合物43b(317mg,0.68mmol)、HMDS(7.2mL,34.3mmol)和MeOH(0.68mL,17.2mmol)为原料,制得红色粉末状固体(43c)287mg,收率91%。1HNMR(600MHz,DMSO-d6)δ11.00(s,1H,imide-NH),7.89(s,1H,Ar-H),7.88(s,1H,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.33(t,2H,J=7.3Hz,Ar-H),7.28(dd,1H,J=8.7Hz,2.3Hz,Ar-H),7.27(t,1H,J=7.8Hz,Ar-H),7.19(d,2H,J=7.3Hz,Ar-H),7.04(t,1H,J=8.2Hz,Ar-H),6.83(dd,1H,J=8.7Hz,5.5Hz,Ar-H),6.76(d,1H,J=8.2Hz,Ar-H),6.63(t,1H,J=7.8Hz,Ar-H),6.53(dt,1H,J=9.2Hz,2.3Hz,Ar-H),5.47(s,2H,-CH2-Ph),4.26(q,2H,J=7.3Hz,-CH2 -CH3),1.34(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.3×2,159.4(d,1JCF=235Hz),137.9,136.5(d,3JCF=12Hz),136.0,133.2,132.2,129.2,129.0×2,128.1,127.5×2,126.8,126.3,123.5,122.7(d,3JCF=11Hz),122.3,121.8,120.1,110.8,108.6(d,2JCF=24Hz),106.3,105.2,97.6(d,2JCF=26Hz),49.9,41.3,15.8.ESI-MSm/z 464.2[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(6-氟-3-吲哚)马来酰亚胺(43)的制备
按照化合物24d的制备方法,以化合物43c(247mg,0.53mmol)、DMSO(0.85mL)、1M的t-BuOK/THF溶液(8.4mL,8.4mmol)及O2为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末固体(43)118mg,收率60%。1H NMR(500MHz,DMSO-d6)δ11.69(s,1H,indole-NH),10.92(s,1H,imide-NH),7.76(s,1H,Ar-H),7.72(s,1H,Ar-H),7.45(d,1H,J=8.2Hz,Ar-H),7.14(d,1H,J=8.6Hz,Ar-H),7.04(t,1H,J=7.9Hz,Ar-H),6.81(d,1H,J=8.1Hz,Ar-H),6.71(dd,1H,J=8.5Hz,5.9Hz,Ar-H),6.67(d,1H,J=7.3Hz,Ar-H),6.47(t,1H,J=8.6Hz,Ar-H),4.24(q,2H,J=7.1Hz,-CH2 -CH3),1.32(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ173.4×2,159.3(d,1JCF=235Hz),136.5(d,3JCF=12Hz),136.0,132.1,130.4,128.2,127.7,126.4,122.6,122.4(d,3JCF=11Hz),122.3,121.8,120.1,110.7,108.2(d,2JCF=24Hz),106.2,105.3,98.4(d,2JCF=27Hz),41.3,15.8.HR-ESIMS m/z 374.1314[M+H]+(calcd.for C22H17N3O2F,374.1305).
化合物44的制备
按照化合物42的合成方法,由化合物43(20mg,0.05mmol)和HCHO(5mL,质量分数37%)合成,以硅胶柱色谱分离、二氯甲烷:甲醇=70:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(6-氟-3-吲哚)马来酰亚胺(44)12mg,收率60%。1H NMR(500MHz,DMSO-d6)δ11.74(s,1H,indole-NH),7.81(s,1H,Ar-H),7.79(d,1H,J=2.7Hz,Ar-H),7.49(d,1H,J=8.3Hz,Ar-H),7.17(dd,1H,J=8.7Hz,2.3Hz,Ar-H),7.06(s,1H,Ar-H),6.84(d,1H,J=8.0Hz,Ar-H),6.73-6.69(m,2H,Ar-H),6.50(td,J=8.4Hz,2.4Hz,1H,Ar-H),6.30(t,1H,J=7.0Hz,-CH2-OH),4.97(d,2H,J=7.0Hz,-CH2 -OH),4.27(q,2H,J=7.2Hz,-CH2 -CH3),1.34(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.6,171.5,159.4(d,1JCF=235Hz),136.62(d,3JCF=12Hz),136.1,132.2×2,130.6,128.2,127.9,127.3,126.4,122.5(d,3JCF=11Hz),121.8,120.3,110.8,108.3(d,2JCF=25Hz),106.1,105.2,98.5(d,2JCF=27Hz),60.8,41.3,15.6.HR-ESIMS m/z 404.1393[M+H]+(calcd.for C23H19N3O3F,404.1405).
化合物45的制备
按照化合物2的制备方法,以化合物43(30mg,0.08mmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(13.5mg,0.16mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-羟甲基-6-氟-3-吲哚)马来酰亚胺(45)35mg,收率99%。1H NMR(600MHz,DMSO-d6)δ7.95(s,1H,Ar-H),7.78(s,1H,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.41(dd,1H,J=9.0Hz,1.5Hz,Ar-H),7.06(t,1H,J=7.7Hz,Ar-H),6.91(d,1H,J=8.1Hz,Ar-H),6.73(t,1H,J=7.9Hz,Ar-H),6.61(dd,J=8.8Hz,5.4Hz,Ar-H),6.51(dd,1H,J=8.8Hz,2.0Hz),5.57(s,2H,-CH2 -OH),4.95(s,2H,-CH2 -OH),4.25(q,2H,J=7.3Hz,-CH2 -CH3),1.31(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.5×2,159.4(d,1JCF=235Hz),136.2(d,3JCF=12Hz),136.1,133.0,132.2,128.3,126.7,126.6,123.2,122.6(d,3JCF=11Hz),122.5,121.8,120.4,110.8,108.8(d,2JCF=25Hz),106.0,105.3,98.0(d,2JCF=26Hz),69.8,60.8,41.3,15.8.HR-ESIMS m/z 456.1342[M+Na]+(calcd.for C24H20N3O4FNa,456.1336).
化合物46的制备
i)1-苄基-6-氯吲哚(46a)的制备
按照化合物24a的合成方法,以6-氯吲哚(303mg,2mmol)、NaH(120mg,3mmol,质量分数60%,分散于石蜡中)和溴化苄(513mg,3mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=60:1(v/v)洗脱得白色粉末状固体(46a)483mg,收率100%。1H NMR(600MHz,CDCl3)δ7.55(d,1H,J=8.22Hz,Ar-H),7.26-7.33(m,4H,Ar-H),7.12(d,1H,J=3.2Hz,Ar-H),7.09(d,2H,J=7.7Hz,Ar-H),7.08(d,1H,J=1.8Hz,Ar-H),6.53(dd,1H,J=3.4Hz,0.9Hz,Ar-H),5.28(s,2H,Ph-CH2-).13C NMR(150MHz,CDCl3)δ137.1,136.8,129.1,129.0,127.9,127.8,127.3,126.8×2,121.9,120.4,109.8,102.0×2,50.2.ESI-MS m/z 242.1/244.1[M+H]+.
ii)2-(1-乙基-3-吲哚)-3-(1-苄基-6-氯-3-吲哚)马来酸酐(46b)的制备
按照化合物24b的合成方法,以化合物46a(419mg,1.73mmol)、(COCl)2(446μL,5.20mmol)、化合物1a(352mg,1.73mmol)和Et3N(480μL,3.47mmol)为原料制备,甲醇重结晶得红色粉末(46b)278mg,收率33.4%。1H NMR(600MHz,DMSO-d6)δ8.01(s,2H,Ar-H),7.59(s,1H,Ar-H),7.54(d,1H,J=8.2Hz,Ar-H),7.34(t,2H,J=7.8Hz,Ar-H),7.29(t,1H,J=7.3Hz,Ar-H),7.19(d,2H,J=7.8Hz,Ar-H),7.11(t,1H,J=7.7Hz,Ar-H),6.92(d,1H,J=8.7Hz,Ar-H),6.77(d,1H,J=8.7Hz,Ar-H),6.75(d,1H,J=7.3Hz,Ar-H),6.69(t,1H,J=7.3Hz,Ar-H),5.53(s,2H,Ph-CH2-),4.32(q,2H,J=7.3Hz,-CH2 -CH3),1.37(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.9,166.8,137.6,136.9,136.3,134.5,133.6,129.9,129.3×2,128.2,127.7,127.5×2,126.8,125.7,125.2,123.2,122.9,122.0,121.0,120.8,111.4,111.2,105.5,104.5,50.0,41.5,15.8.ESI-MS m/z 481.2/483.2[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-6-氯-3-吲哚)马来酰亚胺(46c)的制备
按照按照化合物24c的制备方法,以化合物46b(240mg,0.5mmol)、HMDS(4.2mL,20mmol)和MeOH(0.4mL,10mmol)为原料制备,经硅胶柱色谱分离、二氯甲烷洗脱得红色粉末状固体(46c)218mg,收率91%。1H NMR(600MHz,DMSO-d6)δ11.00(s,1H,imide-NH),7.91(s,1H,Ar-H),7.90(s,1H,Ar-H),7.51(d,1H,J=1.9Hz,Ar-H),7.47(d,1H,J=8.2Hz,Ar-H),7.34(t,2H,J=7.4Hz,Ar-H),7.28(t,1H,J=7.3Hz,Ar-H),7.17(d,2H,J=7.3Hz,Ar-H),7.05(t,1H,J=7.7Hz,Ar-H),6.84(d,1H,J=8.2Hz,Ar-H),6.72(d,1H,J=7.8Hz,Ar-H),6.67(dd,1H,J=8.7Hz,1.9Hz,Ar-H),6.62(t,1H,J=7.3Hz,Ar-H),5.50(s,2H,Ph-CH2-),4.28(q,2H,J=7.3Hz,-CH2 -CH3),1.36(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.3×2,137.9,136.8,136.1,133.5,132.3,129.3,129.2×2,128.1,127.4×2,127.2,126.5,126.2,125.6,122.9,122.3,121.7,120.5,120.2,111.0,110.8,106.3,105.1,49.8,41.3,15.8.ESI-MS m/z 480.1/482.2[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(6-氯-3-吲哚)马来酰亚胺(46)的制备
按照化合物23d的制备方法,以化合物46c(160mg,0.33mmol)、DMSO(0.85mL)、1M的t-BuOK/THF溶液(8.4mL,8.4mmol)及O2为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末(46)128mg,收率99%。1H NMR(500MHz,DMSO-d6)δ11.75(s,1H,indole-NH),10.94(s,1H,imide-NH),7.79(s,1H,Ar-H),7.76(s,1H,Ar-H),7.45(d,1H,J=8.3Hz,Ar-H),7.41(s,1H,Ar-H),7.02(t,1H,J=7.6Hz,Ar-H),6.77(d,1H,J=7.7Hz,Ar-H),6.73(d,1H,J=8.2Hz,Ar-H),6.66(t,1H,J=7.5Hz,Ar-H),6.61(dd,1H,J=8.5Hz,1.1Hz,Ar-H),4.24(q,2H,J=7.2Hz,-CH2 -CH3),1.32(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ173.3×2,136.8,135.9,132.0,130.5,128.4,127.2,126.6,126.2,124.6,122.5,122.2,121.6,120.0,119.9,111.8,110.6,106.1,105.1,41.1,15.7.HR-ESIMS m/z390.1014[M+H]+(calcd.for C22H17N3O2Cl,390.1009).
化合物47的制备
按照化合物42的合成方法,由化合物46(19mg,0.05mmol)和HCHO(5mL,质量分数37%)合成,以硅胶柱色谱分离、二氯甲烷:甲醇=80:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(6-氯-3-吲哚)马来酰亚胺(47)10mg,收率50%。1H NMR(500MHz,DMSO-d6)δ11.78(s,1H,indole-NH),7.83(s,1H,Ar-H),7.79(s,1H,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.42(d,1H,J=1.3Hz,Ar-H),7.04(t,1H,J=7.5Hz,Ar-H),6.77(d,1H,J=8.0Hz,Ar-H),6.71(d,1H,J=8.6Hz,Ar-H),6.68(t,1H,J=7.5Hz,Ar-H),6.62(dd,1H,J=8.6Hz,1.5Hz,Ar-H),6.30(t,1H,J=6.9Hz,imide-CH2-OH),4.94(d,2H,J=6.9Hz,imide-CH2 -OH),4.28(q,2H,J=7.2Hz,-CH2 -CH3),1.33(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.5×2,137.0,136.1,132.3,130.9,128.2,127.0,126.9,126.4,124.6,122.6,122.4,121.7,120.3,120.2,112.1,110.9,106.2,105.2,60.8,41.3,15.8.HR-ESIMS m/z 420.1107[M+H]+(calcd.for C23H19N3O3Cl,420.1109).
化合物48的制备
按照化合物2的制备方法,以化合物46(14mg,36.0μmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(7mg,0.083mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-羟甲基-6-氯-3-吲哚)马来酰亚胺(48)16mg,收率99%。1H NMR(600MHz,DMSO-d6)δ7.99(s,1H,Ar-H),7.83(s,1H,Ar-H),7.69(d,1H,J=1.8Hz,Ar-H),7.50(d,1H,J=8.2Hz,Ar-H),7.06(t,1H,J=7.8Hz,Ar-H),6.89(d,1H,J=8.2Hz,Ar-H),6.75(t,1H,J=7.3Hz,-CH2-OH),6.72(t,1H,J=7.7Hz,Ar-H),6.66(dd,1H,J=8.2Hz,1.8Hz,Ar-H),6.62(d,1H,J=8.2Hz,Ar-H),6.35(t,1H,J=6.8Hz,-CH2-OH),5.61(d,2H,J=7.3Hz,-N-CH2 -OH),4.97(d,2H,J=6.8Hz,-N-CH2 -OH),4.28(q,2H,J=6.8Hz,-CH2 -CH3),1.32(t,3H,J=6.8Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.4×2,136.5,136.0,133.3,132.4,128.6,127.2,126.6,126.3,125.4,122.7,122.5,121.7,120.6,120.4,111.5,110.9,105.9,105.2,69.8,60.9,41.3,15.8.HR-ESIMS m/z 472.1046[M+Na]+(calcd.for C24H20N3O4ClNa,472.1040).
化合物49的制备
i)1-苄基-4-溴吲哚(49a)的制备
按照化合物24a的制备方法,以化合物4-溴吲哚(700mg,3.59mmol),NaH(129mg,5.38mmol,质量分数60%,分散于石蜡中)和溴化苄(0.64mL,5.38mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=100:1(v/v)洗脱得白色晶体(49a)0.88g,收率86%。1H NMR(600MHz,DMSO-d6)δ7.64(d,1H,J=3.3Hz,Ar-H),7.49(d,1H,J=8.3Hz,Ar-H),7.29-7.27(m,2H,Ar-H),7.25-7.23(m,2H,Ar-H),7.19-7.18(m,2H,Ar-H),7.03(t,1H,J=7.7Hz,Ar-H),6.45(d,1H,J=2.8Hz,Ar-H),5.43(s,2H,Ph-CH2-).13C NMR(150MHz,DMSO-d6)δ138.4,136.7,130.9,129.2,129.1,128.0,127.6,127.4,123.0×2,122.4,114.2,110.5,101.4,50.0.ESI-MS m/z 286.0/288.0[M+H]+.
ii)2-(1-乙基-3-吲哚)-3-(1-苄基-4-溴-3-吲哚)马来酸酐(49b)的制备
按照化合物24b的制备方法,以化合物49a(400mg,1.4mmol)、(COCl)2(214μL,2.25mmol)、化合物1a(548mg,2.7mmol)和Et3N(626μL,4.5mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=7:1(v/v)洗脱得红色粉末(49b)220mg,收率30.0%。1H NMR(600MHz,pyridine-d5)δ8.37(s,1H,Ar-H),7.63(s,1H,Ar-H),7.41(d,1H,J=3.8Hz,Ar-H),7.42(m,3H,Ar-H),7.24-7,22(m,4H,Ar-H),7.06-7.04(m,4H,Ar-H),6.77(t,1H,J=7.2Hz,Ar-H),5.3(d,1H,J=15.9Hz,Ph-CH2 -),5.24(d,1H,J=15.9Hz,Ph-CH2-),4.05(q,2H,J=8.2Hz,-CH2 -CH3),1.37(t,3H,J=8.2Hz,-CH2-CH3 ).13C NMR(150MHz,pyridine–d5)δ166.0,165.6,148.5×2,136.4,135.5,135.4,133.2,130.6,127.5,126.4,125.5,125.4,124.5,124.1,123.8,122.5,122.1,121.5,121.2,119.9,113.4,109.3,109.2,104.4,103.8,48.8,40.1,13.5.ESI-MS m/z 525.1/527.1[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-4-溴-3-吲哚)马来酰亚胺(49c)的制备
按照化合物24c的制备方法,以化合物49c(110mg,0.21mmol)、HMDS(4.5mL,21mmol)和MeOH(0.4mL,10.5mmol)为原料制备,硅胶柱色谱分离、石油醚:二氯甲烷=1:4(v/v)洗脱得红色粉末(49c)100mg,收率91%。1H NMR(600MHz,DMSO-d6)δ11.02(s,1H,imide-NH),8.05(s,1H,Ar-H),7.53(s,1H,Ar-H),7.47(t,2H,J=8.7Hz,Ar-H),7.27(d,1H,J=7.6Hz,Ar-H),7.19(dd,3H,J=5.0Hz,1.8Hz,Ar-H),7.09(dt,2H,J=8.2Hz,2.0Hz,Ar-H),6.99-6.95(m,2H,Ar-H),6.68(d,1H,J=8.1Hz,Ar-H),6.58(t,1H,J=7.6Hz,Ar-H),5.42(d,1H,J=15.9Hz)/5.38(d,1H,J=15.9Hz)(Ph-CH2-),4.25(q,2H,J=7.2Hz,-CH2 -CH3),1.35(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.9,173.2,137.9,137.6,136.5,134.3,133.3,132.0,129.0,127.9,127.1,126.9,126.7,125.7,124.6,123.7,122.5,121.9,120.6,114.4,111.0×2,110.9,106.2,105.0,49.8,49.2,41.4,15.8.ESI-MS m/z 524.1/526.1[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(4-溴-3-吲哚)马来酰亚胺(49)的制备
按照化合物24d的制备方法,以化合物49c(110mg,0.21mmol)、DMSO(1.24mL)、1M的t-BuOK/THF溶液(0.97mL,0.97mmol)及O2为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色粉末(49)63mg,收率70%。1H NMR(600MHz,DMSO-d6)δ11.65(s,1H,indole-NH),10.93(s,1H,imide-NH),8.00(s,1H,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.45(d,1H,J=8.3Hz,Ar-H),7.30(d,1H,J=2.8Hz,Ar-H),7.24(d,1H,J=7.7Hz,Ar-H),7.09(t,1H,J=7.7Hz,Ar-H),7.01(t,1H,J=6.6Hz,Ar-H),6.58(d,1H,J=7.9Hz,Ar-H),6.53(t,1H,J=7.3Hz,Ar-H),4.24(q,2H,J=7.2Hz,-CH2 -CH3),1.34(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ174.0,173.4,137.9,136.4,134.3,133.3,128.6,127.4,126.4,125.8,124.1,123.5,122.4,121.8,120.4,114.1,112.2,110.8,106.5,105.1,41.3,15.8.HR-ESIMS m/z 434.0514[M+H]+(calcd.for C22H17N3O2Br,434.0504).
化合物50的制备
按照化合物42的合成方法,由化合物49(20mg,0.05mmol)和HCHO(5mL,质量分数37%)合成,以硅胶柱色谱分离、二氯甲烷:甲醇=70:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(4-溴-3-吲哚)马来酰亚胺(50)11mg,收率50%。1H NMR(500MHz,DMSO-d6)δ11.70(s,1H,indole-NH),8.03(s,1H,Ar-H),7.48(d,1H,J=8.1Hz,Ar-H),7.45(d,1H,J=8.2Hz,Ar-H),7.34(d,1H,J=2.5Hz,Ar-H),7.22(d,1H,J=7.5Hz,Ar-H),7.07(t,1H,J=7.9Hz,Ar-H),7.02(t,1H,J=7.3Hz,Ar-H),6.56(d,2H,J=7.5Hz,Ar-H),6.34(t,1H,J=7.0Hz,-CH2-OH),4.95(d,2H,J=7.0Hz,-CH2 -OH),4.27(q,2H,J=7.2Hz,-CH2 -CH3),1.33(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.9,171.4,137.8,136.4,133.9,133.3,128.6,126.7,126.2,125.6,124.0,123.4,122.4,121.6,120.4,113.8,112.1,110.8,106.1,105.0,60.7,41.2,15.6.HR-ESIMS m/z 464.0602[M+H]+(calcd.for C23H19N3O3Br,464.0604).
化合物51的制备
按照化合物2的制备方法,以化合物49(100mg,0.23mmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(97mg,1.15mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱分离得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-羟甲基-4-溴-3-吲哚)马来酰亚胺(51)67mg,收率59%。1H NMR(500MHz,DMSO-d6)δ8.02(s,1H,Ar-H),7.68(d,1H,J=8.1Hz,Ar-H),7.48(s,1H,Ar-H),7.45(d,1H,J=8.3Hz,Ar-H),7.28(d,1H,J=7.5Hz,Ar-H),7.15(d,1H,J=7.9Hz,Ar-H),7.06(t,1H,J=7.4Hz,Ar-H),6.73(d,1H,J=8.2Hz,Ar-H),6.62(t,1H,J=6.8Hz,Ar-H),6.60(t,1H,J=7.2Hz,indole-CH2-OH),6.37(t,1H,J=6.5Hz,imide-CH2-OH),5.53(d,2H,J=7.2Hz,indole-CH2 -OH),4.97(d,2H,J=6.5Hz,imide-CH2 -OH),4.24(q,2H,J=7.1Hz,-CH2 -CH3),1.33(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.0,171.5,137.4,136.5,134.6,133.5,131.3,127.2,126.2,125.9,124.8,123.8,122.6,121.9,120.7,114.1,111.2,110.9,106.0,105.1,69.6,60.8,41.4,15.7.HR-ESIMS m/z516.0540[M+Na]+(calcd.for C24H20N3O4BrNa,516.0535).
化合物52的制备
i)1-苄基-5-溴吲哚(52a)的制备
按照化合物24a的制备方法,以化合物5-溴吲哚(700mg,3.59mmol)、NaH(129mg,5.38mmol,质量分数60%,分散于石蜡中)和溴化苄(0.64mL,5.38mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=60:1(v/v)洗脱得白色结晶粉末(52a)0.88g,收率86%。1HNMR(600MHz,DMSO-d6)δ7.75(d,1H,J=1.6Hz,Ar-H),δ7.56(d,1H,J=3.3Hz,Ar-H),7.42(d,2H,J=8.8Hz,Ar-H),7.30(t,2H,J=7.7Hz,Ar-H),7.24-7.16(m,3H,Ar-H),6.48(d,1H,J=2.2Hz,Ar-H),5.41(s,2H,-CH2-).13C NMR(150MHz,DMSO-d6)δ138.5,135.0,131.3,130.7×2,129.1,128.0,127.6,127.5,124.2,123.2,112.8,112.4,101.3,49.8.ESI-MS m/z 286.0/288.0[M+H]+.
ii)2-(1-乙基-3-吲哚)-3-(1-苄基-5-溴-3-吲哚)马来酸酐(52b)的制备
按照化合物24b的制备方法,以化合物52a(1100mg,3.86mmol)、(COCl)2(500μL,5.79mmol)、化合物1a(783mg,3.86mmol)和Et3N(1.07mL,7.72mmol)为原料制备,纯甲醇重结晶得红色粉末(52b)652mg,收率32.2%。1H NMR(600MHz,DMSO-d6)δ8.06(s,1H,Ar-H),7.99(s,1H,Ar-H),7.55(d,1H,J=8.1Hz,Ar-H),7.39(d,1H,J=8.7Hz,Ar-H),7.33(t,2H,J=7.5Hz,Ar-H),7.28(d,1H,J=6.7Hz,Ar-H),7.17(d,2H,J=7.4Hz,Ar-H),7.14(d,1H,J=8.8Hz,Ar-H),7.10(t,1H,J=7.7Hz,Ar-H),6.98(s,1H,Ar-H),6.76(d,1H,J=8.1Hz,Ar-H),6.69(d,1H,J=8.2Hz,Ar-H),5.52(s,2H,-CH2-Ph),4.33(q,2H,J=6.6Hz,-CH2 -CH3),1.39(t,3H,J=6.6Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.8×2,137.6,136.2,135.2,134.8,133.2,129.5,129.2,128.2,128.0,127.6,127.5,127.4,125.8,125.3,124.4,122.9,121.7,120.7,113.5,113.4,111.2,104.8,104.6,99.9,50.2,41.5,16.1.ESI-MS m/z 525.1/527.0[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-5-溴-3-吲哚)马来酰亚胺(52c)的制备
按照化合物24c的制备方法,以化合物52b(110mg,0.21mmol)、HMDS(4.5mL,21mmol)和MeOH(0.4mL,11mmol)为原料制备,经硅胶柱色谱分离、二氯甲烷洗脱得红色粉末状固体(52c)100mg,收率91%。1H NMR(600MHz,DMSO-d6)δ10.98(s,1H,imide-NH),7.96(s,1H,Ar-H),7.86(s,1H,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.34(dt,3H,J=7.2Hz,1.7Hz,Ar-H),7.27(t,1H,J=7.3Hz,Ar-H),7.14(d,2H,J=7.1Hz,2H,Ar-H),7.06(dd,1H,J=6.8Hz,2.0Hz,Ar-H),6.90(d,1H,J=1.9Hz,Ar-H),6.72(d,1H,J=8.0Hz,Ar-H),6.62(t,1H,J=7.2Hz,Ar-H),5.49(s,2H,-CH2-Ph),4.30(q,2H,J=7.2Hz,-CH2 -CH3),1.38(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.2×2,137.9,136.0,135.0,133.9,132.0,129.2,128.8,128.4,128.19,127.4,127.0×2,126.7,124.8,124.2×2,122.4,121.4,120.1,113.1,112.9,110.8,105.6,105.1,50.0,41.3,16.1.ESI-MS m/z 524.1/526.1[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(5-溴-3-吲哚)马来酰亚胺(52)的制备
按照化合物24d的制备方法,以化合物52c(90mg,0.172mmol)、DMSO(0.85mL)和1M的t-BuOK/THF溶液(7mL,7mmol)及O2为原料制备。硅胶柱色谱分离、二氯甲烷:乙酸乙酯=4:1(v/v)洗脱得红色粉末(52)54mg,收率73%。1H NMR(600MHz,DMSO-d6)δ11.82(s,1H,indole-NH),10.92(s,1H,imide-NH),7.80(s,1H,Ar-H),7.77(s,1H,Ar-H),7.49(d,1H,J=8.3Hz,Ar-H),7.31(d,1H,J=8.6Hz,Ar-H),7.05(d,1H,J=8.0Hz,Ar-H),7.03(d,1H,J=8.0Hz,Ar-H),6.79(s,1H,Ar-H),6.77(d,1H,J=8.0Hz,Ar-H),6.67(t,1H,J=7.2Hz,Ar-H),4.28(q,2H,J=7.2Hz,-CH2 -CH3),1.36(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.4,173.3,136.0,135.2,131.9,131.0,128.1,127.8,127.5,126.5,124.6,124.0,122.4,121.4,120.1,114.2,112.4,110.7,105.8,105.3,41.3,16.1.HR-ESIMS m/z434.0514[M+H]+(calcd.for C22H17N3O2Br,434.0504).
化合物53的制备
按照化合物42的合成方法,由化合物52(21mg,0.05mmol)和HCHO(5mL,质量分数37%)合成,以硅胶柱色谱分离、二氯甲烷:甲醇=70:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(5-溴-3-吲哚)马来酰亚胺(53)12mg,收率55%。1H NMR(500MHz,DMSO-d6)δ11.86(s,1H,indole-NH),7.83(s,1H,Ar-H),7.80(s,1H,Ar-H),7.49(d,1H,J=8.4Hz,Ar-H),7.32(d,1H,J=8.7Hz,Ar-H),7.06(d,1H,J=8.3Hz,Ar-H),7.03(d,1H,J=7.7Hz,Ar-H),6.81(s,1H,Ar-H),6.77(t,1H,J=8.2Hz,Ar-H),6.67(t,1H,J=7.7Hz,Ar-H),6.31(t,1H,J=6.7Hz,-CH2-OH),4.95(d,2H,J=6.4Hz,-CH2 -OH),4.28(q,2H,J=7.0Hz,-CH2 -CH3),1.36(t,3H,J=6.8Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.4,171.3,136.0,135.1,131.9,131.1,127.6,127.3,127.2,126.2,124.6,123.9,122.3,121.3,120.1,114.1,112.4,110.7,105.5,105.0,60.7,41.2,15.9.HR-ESIMS m/z464.0600[M+H]+(calcd.for C23H19N3O3Br,464.0604).
化合物54的制备
按照化合物2的制备方法,以化合物53(53mg,0.122mmol)、NaHCO3(51mg,0.62mmol)和甲醛溶液(3mL,质量分数37%)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-羟甲基-5-溴-3-吲哚)马来酰亚胺(54)60mg,收率99%。1H NMR(500MHz,DMSO-d6)δ8.04(s,1H,Ar-H),7.78(s,1H,Ar-H),7.50(d,2H,J=8.6Hz,Ar-H),7.13(d,1H,J=8.6Hz,Ar-H),7.06(t,1H,J=7.5Hz,Ar-H),6.87(d,1H,J=7.9Hz,Ar-H),6.75(t,1H,J=6.8Hz,indole-CH2-OH),6.71(d,1H,J=7.6Hz,Ar-H),6.68(s,1H,Ar-H),6.34(t,1H,J=6.4Hz,imide-CH2-OH),5.58(d,2H,J=6.8Hz,indole-CH2 -OH),4.96(d,2H,J=6.4Hz,imide-CH2 -OH),4.28(q,2H,J=6.9Hz,-CH2 -CH3),1.35(t,3H,J=6.9Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.4×2,136.0,134.8,133.6,132.0,128.2,128.1,126.9,126.6,124.8,124.3,122.5,121.4,120.4,113.3,113.2,110.8,105.4,105.2,69.9,60.8,41.4,16.1.HR-ESIMS m/z 516.0547[M+Na]+(calcd.for C24H20N3O4BrNa,516.0535),.
化合物55(化合物26d)的制备
见化合物26d的制备。
化合物56的制备
按照化合物42的合成方法,由化合物55(21mg,0.05mmol)和HCHO(5mL,质量分数37%)合成,以硅胶柱色谱分离、二氯甲烷:甲醇=70:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(56)12mg,收率53%。1H NMR(600MHz,DMSO-d6)δ11.81(s,1H,indole-NH),7.85(s,1H,Ar-H),7.80(s,1H,Ar-H),7.58(d,1H,J=1.8Hz,Ar-H),7.50(d,1H,J=8.3Hz,Ar-H),7.38(dd,1H,J=7.6Hz,1.5Hz,Ar-H),7.06(t,1H,J=7.6Hz,Ar-H),6.79(d,1H,J=8.0Hz,Ar-H),6.76(dd,1H,J=8.6Hz,1.8Hz,Ar-H),6.71(d,1H,J=3.9Hz,Ar-H),6.69(t,1H,J=3.9Hz,Ar-H),6.32(t,1H,J=7.0Hz,-CH2-OH),4.96(d,2H,J=7.0Hz,-CH2 -OH),4.29(q,2H,J=7.2Hz,-CH2 -CH3),1.35(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.5×2,137.5,136.1,132.3,130.7,128.2,127.0,126.3,124.9,123.0,122.8,122.4,121.7,120.3,115.1,115.0,110.9,106.2,105.1,60.8,41.3,15.8.HR-ESIMS m/z464.0591[M+H]+(calcd.for C23H19N3O3Br,464.0604).
化合物57的制备
按照化合物2的制备方法,以化合物56(50mg,0.115mmol)、NaHCO3(29mg,0.345mmol)和甲醛溶液(3mL,质量分数37%)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-羟甲基-6-溴-3-吲哚)马来酰亚胺(57)48mg,收率85%。1H NMR(500MHz,DMSO-d6)δ7.98(s,1H,Ar-H),7.83(s,2H,Ar-H),7.49(d,1H,J=8.2Hz,Ar-H),7.07(t,1H,J=7.7Hz,Ar-H),6.89(d,1H,J=7.7Hz,Ar-H),6.78(d,1H,J=8.2Hz,Ar-H),6.76(t,1H,J=6.8Hz,-CH2-OH),6.72(t,1H,J=7.7Hz,Ar-H),6.60(d,1H,J=8.2Hz,Ar-H),6.35(t,1H,J=6.6Hz,-CH2-OH),5.61(d,2H,J=6.6Hz,-N-CH2 -OH),4.98(d,2H,J=6.6Hz,-CH2 -OH),4.27(q,2H,J=6.6Hz,-CH2 -CH3),1.34(t,3H,J=6.6Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.4×2,136.9,136.1,133.1,132.3,128.7,126.6,126.3,125.7,123.2,123.1,122.5,121.7,120.4,115.3,114.4,110.9,106.0,105.2,69.8,60.9,41.4,15.8.HR-ESIMS m/z 516.0544[M+Na]+(calcd.for C24H20N3O4BrNa,516.0535).
化合物58的制备
i)1-苄基-7-溴吲哚(58a)的制备
按照化合物24a的制备方法,以化合物7-溴吲哚(700mg,3.59mmol)、NaH(129mg,5.38mmol,质量分数60%,分散于石蜡中)和溴化苄(0.64mL,5.38mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=60:1(v/v)洗脱得白色结晶粉末(58a)1.0g,收率98%。1HNMR(600MHz,DMSO-d6)δ7.61(d,1H,J=7.8Hz,Ar-H),7.54(d,1H,J=3.2Hz,Ar-H),7.29(d,1H,J=7.5Hz,Ar-H),7.26(t,2H,J=7.5Hz,Ar-H),7.21(d,1H,J=7.1Hz,Ar-H),6.95(d,1H,J=7.7Hz,Ar-H),6.93(dd,2H,J=7.6Hz,2.7Hz,Ar-H),6.61(d,1H,J=3.2Hz,Ar-H),5.81(s,2H,-CH2-).13C NMR(150MHz,DMSO-d6)δ140.2×2,133.3,132.4,129.1×2,127.6,127.0,126.3,121.4×2,121.1,103.6,102.6,51.0.ESI-MS m/z 286.0/288.0[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-7-溴-3-吲哚)马来酸酐(58b)的制备
按照化合物24b的制备方法,以化合物58a(700mg,2.5mmol)、(COCl)2(714μL,7.5mmol)、化合物1a(686mg,3.3mmol)和Et3N(0.723mL,5.2mmol)为原料制备,纯甲醇重结晶得红色粉末(58b)380mg,收率30%。1H NMR(600MHz,DMSO-d6)δ8.04(s,1H,Ar-H),8.01(s,1H,Ar-H),7.50(d,1H,J=8.0Hz,Ar-H),7.31(t,2H,J=7.0Hz,Ar-H),7.25(t,1H,J=7.4Hz,Ar-H),7.21(d,1H,J=7.6Hz,Ar-H),7.09(t,1H,J=7.6Hz,Ar-H),6.99(d,2H,J=7.4Hz,Ar-H),6.94(d,1H,J=8.0Hz,Ar-H),6.77(d,1H,J=8.0Hz,Ar-H),6.71(t,1H,J=7.5Hz,Ar-H),6.63(t,1H,J=7.8Hz,Ar-H),5.90(s,2H,-CH2-Ph),4.31(q,2H,J=5.8Hz,-CH2 -CH3),1.34(t,3H,J=5.8Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.8,166.7,139.4,136.4,136.3,133.8,132.6,131.1,129.8,129.2,128.2,127.8,127.3,126.2,125.9,125.8,123.0,121.8,121.4,120.9,120.8,119.4,111.2,105.8,104.6,103.9,51.7,41.6,15.8.ESI-MS m/z 525.1/527.0[M+H]+.
iii)2-(1-乙基-3-吲哚)-3-(1-苄基-7-溴-3-吲哚)马来酰亚胺(58c)的制备
按照化合物24c的制备方法,以化合物58b(110mg,0.45mmol)、HMDS(4.5mL,21mmol)和MeOH(0.4mL,11mmol)为原料制备,经硅胶柱色谱分离、二氯甲烷洗脱得红色粉末状固体(58c)100mg,收率91%。1H NMR(600MHz,DMSO-d6)11.04(s,1H,imide-NH),7.93(s,1H,Ar-H),7.92(s,1H,Ar-H),7.45(d,1H,J=8.3Hz,Ar-H),7.31(t,2H,J=7.5Hz,Ar-H),7.25(s,1H,J=7.4Hz,Ar-H),7.15(dd,1H,J=7.6Hz,1.0Hz,Ar-H),7.04(d,1H,J=1.1Hz,Ar-H),6.97(d,2H,J=7.2Hz,Ar-H),6.88(dd,1H,J=8.0Hz,1.0Hz,Ar-H),6.76(d,1H,J=8.0Hz,Ar-H),6.67(d,1H,J=7.9Hz,Ar-H),6.56(t,1H,J=7.8Hz,Ar-H),5.88(s,2H,-CH2-Ph),4.27(q,2H,J=7.2Hz,-CH2 -CH3),1.35(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.2×2,139.7,136.0,135.6,135.5,132.5,132.4,130.3,130.2,129.1,127.7×2,126.3,126.2,125.7,122.4,121.7,121.6,121.1×2,120.2,110.8,106.5,105.1,103.7,51.5,41.4,15.8.ESI-MS m/z 523.8/525.8[M+H]+.
iv)2-(1-乙基-3-吲哚)-3-(7-溴-3-吲哚)马来酰亚胺(58)的制备
按照化合物24d的制备方法,以化合物58c(292mg,0.558mmol)、DMSO(0.85mL)和1M的t-BuOK/THF溶液(8.4mL,8.4mmol)及O2为原料制备。硅胶柱色谱分离、二氯甲烷:乙酸乙酯=4:1(v/v)洗脱得红色粉末(58)150mg,收率64%。1H NMR(500MHz,DMSO-d6)δ11.86(s,1H,indole-NH),10.96(s,1H,imide-NH),7.78(s,1H,Ar-H),7.72(s,1H,Ar-H),7.46(d,1H,J=8.2Hz,Ar-H),7.19(d,1H,J=7.5Hz,Ar-H),7.03(t,1H,J=7.6Hz,Ar-H),6.82(d,1H,J=8.0Hz,Ar-H),6.76(d,1H,J=8.0Hz,Ar-H),6.69(t,1H,J=7.5Hz,Ar-H),6.57(t,1H,J=7.7Hz,Ar-H),4.24(q,2H,J=7.1Hz,-CH2 -CH3),1.31(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ173.1×2,135.9,134.7,132.1,130.3,128.9,127.4,127.1,126.2,124.6,122.2,121.5,121.1,120.8,120.1,110.6,107.2,105.1,104.6,41.1,15.7.HR-ESIMS m/z 434.0509[M+H]+(calcd.for C22H17N3O2Br,434.0504).
化合物59的制备
按照化合物42的合成方法,由化合物58(21mg,0.05mmol)和HCHO(5mL,质量分数37%)合成,以硅胶柱色谱分离、二氯甲烷:甲醇=70:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(7-溴-3-吲哚)马来酰亚胺(59)9mg,收率40%。1H NMR(500MHz,DMSO-d6)δ11.94(s,1H,indole-NH),7.84(s,1H,Ar-H),7.79(s,1H,Ar-H),7.49(d,1H,J=8.3Hz,Ar-H),7.22(d,1H,J=7.5Hz,Ar-H),7.06(t,1H,J=7.6Hz,Ar-H),6.86(d,1H,J=8.1Hz,Ar-H),6.77(d,1H,J=8.0Hz,Ar-H),6.73(d,1H,J=7.6Hz,Ar-H),6.59(t,1H,J=7.8Hz,Ar-H),6.34(t,1H,J=7.0Hz,N-CH2-OH),4.97(d,2H,J=7.0Hz,N-CH2 -OH),4.27(q,2H,J=7.2Hz,-CH2 -CH3),1.33(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.5×2,136.1,134.9,132.4,130.6,128.7,127.4,126.8,126.3,124.9,122.5,121.7,121.4,120.9,120.4,110.9,107.2,105.1,104.9,60.8,41.3,15.8.HR-ESIMS m/z 464.0592[M+H]+(calcd.for C23H19N3O3Br,464.0604).
化合物60的制备
按照化合物2的制备方法,以化合物59(75mg,0.173mmol)、NaHCO3(73mg,0.866mmol)和甲醛溶液(3mL,质量分数37%)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-羟甲基-7-溴-3-吲哚)马来酰亚胺(60)20mg,收率25%。1H NMR(600MHz,CDCl3)δ7.64(s,1H,Ar-H),7.61(s,1H,Ar-H),7.25(d,1H,J=6.6Hz,Ar-H),7.21(d,1H,J=7.6Hz,Ar-H),7.07(t,1H,J=7.6Hz,Ar-H),6.90(d,1H,J=8.2Hz,Ar-H),6.77(d,1H,J=8.0Hz,Ar-H),6.74(t,1H,J=7.9Hz,Ar-H),6.50(t,1H,J=7.9Hz,Ar-H),5.86(d,2H,J=7.2Hz,-CH2 OH),5.14(d,2H,J=6.1Hz,-CH2 OH),4.26(t,1H,J=7.2Hz,-CH2OH),4.17(t,1H,J=6.1Hz,-CH2OH),4.09(q,2H,J=7.3Hz,-CH2 -CH3),1.38(t,3H,J=7.3Hz,-CH2-CH3 ).13C NMR(150MHz,CDCl3)δ172.0,171.7,136.0,134.1,132.3,132.0,129.9,129.7,127.7,126.3,124.6,122.5,122.2,121.7,121.6,120.6,109.7,106.7,105.5,103.5,71.1,61.5,41.6,15.2.HR-ESIMS m/z 516.0540[M+Na]+(calcd.for C24H20N3O4BrNa,516.0535).
化合物61的制备
以6mL DMF溶解化合物55(300mg,0.693mmol)于50ml单口瓶中,加入PPh3(36.3mg,0.139mmol)、PdCl2(6.1mg,0.0346mmol)和丙烯基三丁基锡(258μL,0.83mmol),氩气保护下,110℃油浴搅拌反应23h。冷却至室温,***萃取,使用饱和NaCl水溶液洗涤3~4次,以无水硫酸钠干燥有机相,真空旋蒸至干。抽滤除去PdCl2,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得到红色固体2-(1-乙基-3-吲哚)-3-(6-烯丙基-3-吲哚)马来酰亚胺(61)171mg,收率62%。1H NMR(600MHz,DMSO-d6)δ11.60(s,1H,indole-NH),10.90(s,1H,imide-NH),7.70(d,1H,J=2.9Hz,Ar-H),7.70(s,1H,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.17(s,1H,Ar-H),7.05(t,1H,J=7.1Hz,Ar-H),6.92(d,1H,J=8.1Hz,Ar-H),6.71(t,1H,J=7.5Hz,Ar-H),6.64(d,1H,J=8.1Hz,Ar-H),6.47(d,1H,J=8.3Hz,Ar-H),5.94-5.86(m,1H,ArCH2CH=CH2),5.00-4.97(m,1H,ArCH2CH=CH2),4.96-4.94(m,1H,ArCH2CH=CH2 ),4.23(q,2H,J=7.3Hz,-CH2 -CH3),3.32(d,2H,J=6.7Hz,Ar-CH2 CH=CH2),1.30(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ173.6,173.5,138.8,137.0,136.0,133.7,131.9,129.6,128.6,127.4,126.6,124.1,122.2,121.9,121.5,121.1,120.1,115.9,111.8,110.6,106.0,105.5,41.2,39.0,15.8.HR-ESIMS m/z 396.1718[M+H]+(calcd.forC25H22N3O2,396.1712).
化合物62的制备
以20mL CH2Cl2溶解化合物61(128mg,0.324mmol)于50ml单口烧瓶中,40℃下冷凝水回流0.5h,使化合物全部溶解,冷却至室温,加入2-甲基-2-丁烯(1.2mL,11.34mmol)和Grubbs,2代催化剂(27.5mg,0.0324mmol),氩气保护下,40℃下冷凝水回流2h,冷却至室温,真空旋蒸至干。硅胶柱色谱分离、石油醚:乙酸乙酯=6:1(v/v)洗脱得红色固体2-(1-乙基-3-吲哚)-3-(6-异戊烯基-3-吲哚)马来酰亚胺(62)91mg,收率66%。1H NMR(600MHz,DMSO-d6)δ11.50(s,1H,indole-NH),10.87(s,1H,imide-NH),7.67(d,1H,J=1.7Hz,Ar-H),7.66(s,1H,Ar-H),7.47(d,1H,J=8.3Hz,Ar-H),7.14(s,1H,Ar-H),7.06(t,1H,J=7.1Hz,Ar-H),6.96(d,1H,J=8.0Hz,Ar-H),6.74(t,1H,J=7.2Hz,Ar-H),6.64(d,1H,J=8.2Hz,Ar-H),6.47(dd,1H,J=8.3Hz,1.4Hz,Ar-H),5.27-5.22(m,1H,ArCH2CH=C(CH3)2),4.22(q,2H,J=7.2Hz,-CH2 -CH3),3.27(d,2H,J=7.4Hz,ArCH2 CH=C(CH3)2),1.67(s,3H,ArCH2CH=C(CH3 )2),1.64(s,3H,ArCH2CH=C(CH3 )2),1.29(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ172.9,172.8,136.4,135.4,134.8,131.2,131.1,128.9,128.0,126.7,125.9,123.8,123.1,121.6,121.3,120.8,120.3,119.4,101.0,105.3,104.8,100.0,40.5,33.6,25.4,17.5,15.0.HR-ESIMS m/z 424.2029[M+H]+(calcd.for C27H26N3O2,424.2025).
化合物63的制备
按照化合物2的制备方法,以化合物62(43mg,0.1088mmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(46mg,0.544mmol)为原料制备,得深红色固体N-羟甲基-2-(1-乙基-3-吲哚)-3-(1-羟甲基-6-异戊烯基-3-吲哚)马来酰亚胺(63)17mg,收率34%。1H NMR(600MHz,DMSO-d6)δ7.90(s,1H,Ar-H),7.63(s,1H,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.34(s,1H,Ar-H),7.10(d,1H,J=7.9Hz,Ar-H),7.08(d,1H,J=8.3Hz,Ar-H),6.79(t,1H,J=7.0Hz,-CH2OH),6.73(t,1H,J=7.2Hz,Ar-H),6.48(q,2H,J=8.2Hz,Ar-H),6.38(t,1H,J=6.9Hz,imide-CH2OH),5.55(d,2H,J=7.0Hz,indole-CH2 OH),5.23(t,1H,J=7.2Hz,ArCH2CH=C(CH3)2),4.96(d,2H,J=6.9Hz,-CH2 OH),4.22(q,2H,J=7.2Hz,-CH2 -CH3),3.28(d,2H,J=7.4Hz,ArCH2 CH=C(CH3)2),1.65(s,3H,ArCH2CH=C(CH3 )2),1.63(s,3H,ArCH2CH=C(CH3 )2),1.26(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ171.6×2,136.6,136.0,135.9,132.4,132.0,131.8,127.7,127.5,126.8,124.4,122.4,121.9,121.7,121.5,120.4,110.8,110.6,105.6,105.4,100.0,69.5,60.8,41.2,34.5,26.1,18.2,15.6.ESI-MSm/z 484.2[M+H]+.
化合物64的制备
在敞口石英瓶中,用1.0L丙酮溶解化合物1(40mg,0.1mmol),加催化量的I2,在250W汞灯下照射搅拌24h,真空蒸去大部分溶剂后,倒入100mL Na2S2O3饱和水溶液中,搅拌10min,乙酸乙酯萃取(50mL×3次),合并有有机层,并用无水Na2SO4干燥,真空蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色荧光粉末12-乙基-13-氰甲基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(64)28mg,收率71%。1H NMR(600MHz,DMSO-d6)δ11.27(s,1H,indole-NH),9.12(d,1H,J=8.3Hz,Ar-H),9.11(d,1H,J=8.3Hz,Ar-H),7.96(d,1H,J=8.2Hz,Ar-H),7.91(d,1H,J=8.2Hz,Ar-H),7.73(t,1H,J=7.7Hz,Ar-H),7.67(t,1H,J=7.9Hz,Ar-H),7.53(t,1H,J=7.3Hz,Ar-H),7.43(t,1H,J=7.3Hz,Ar-H),5.79(s,2H,-N-CH 2-CN),4.75(q,2H,J=6.8Hz,-CH 2-CH3),1.10(t,3H,J=6.8Hz,-CH2-CH 3).13C NMR(150MHz,DMSO-d6)δ171.0,170.9,152.0,145.5,144.4,133.1,132.9,128.8,128.5,125.5,125.4,124.5,123.6,123.5,123.0,122.1,121.5,121.0,116.4,113.3,113.0,43.3,31.0,14.0.HR-ESIMS m/z 391.1206[M–H]–(calcd.forC24H15N4O2,391.1195).
化合物65的制备
按照化合物2的制备方法,以化合物2(10mg,24.6μmol)和NaHCO3(4.1mg,49.2μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色荧光粉末6-羟甲基-12-乙基-13-氰乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(65)10.1mg,收率94%。1H NMR(600MHz,CDCl3)δ9.24(d,1H,J=7.8Hz,Ar-H),9.22(d,1H,J=7.8Hz,Ar-H),7.66(dt,1H,J=7.5Hz,1.4Hz,Ar-H),7.65(d,1H,J=6.9Hz,Ar-H),7.64(dt,1H,J=7.8Hz,0.9Hz,Ar-H),7.59(d,1H,J=7.8Hz,Ar-H),7.49(dt,1H,J=7.8Hz,0.9Hz,Ar-H),7.46(dt,1H,J=7.5Hz,1.4Hz,Ar-H),5.40(d,2H,J=7.8Hz,-N-CH 2-OH),4.96(t,2H,J=7.8Hz,N-CH 2-CH2CN),4.66(q,2H,J=7.3Hz,-CH 2-CH3),3.18(t,1H,J=7.8Hz,NCH2-OH),2.21(t,2H,J=7.8Hz,NCH2-CH 2-CN),1.12(t,3H,J=7.3Hz,-CH3).13C NMR(150MHz,CDCl3)δ169.0,168.9,145.0,143.8,133.9,132.7,128.9,128.4,128.3,126.5,126.1,125.4,124.5,123.3,122.5,122.4,121.5,120.2,116.4,112.2,111.7,61.8,44.4,43.9,15.7,13.6.ESI-MS m/z 437.2[M+H]+.
化合物66的制备
按照化合物64的制备方法,以化合物3(30mg,0.072mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色荧光粉末12-乙基-13-氰丙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(66)20.6mg,收率68%。1H NMR(600MHz,CDCl3)δ9.25(d,1H,J=8.2Hz,Ar-H),9.24(d,1H,J=8.3Hz,Ar-H),7.77(s,1H,imide-NH),7.64(t,1H,J=7.4Hz,Ar-H),7.63(d,1H,J=7.4Hz,Ar-H),7.62(t,1H,J=7.7Hz,Ar-H),7.60(d,1H,J=7.8Hz,Ar-H),7.48(dt,1H,J=6.8Hz,1.7Hz,Ar-H),7.46(dt,1H,J=7.4Hz,1.7Hz,Ar-H),4.84(t,2H,J=7.1Hz,N-CH 2-(CH2)2CN),4.68(q,2H,J=7.1Hz,N-CH 2-CH3),1.77(t,2H,J=6.9Hz,N(CH2)2-CH 2-CN),1.26-1.24(m,2H,NCH2-CH 2-CH2CN),1.10(t,3H,J=7.1Hz,-CH2-CH 3).13C NMR(150MHz,CDCl3)δ169.5×2,145.0,144.1,133.7,133.4,128.1,128.0,126.4,126.1,124.6,124.5,122.6,122.3,122.1,121.6,121.5,121.2,118.2,112.2,111.8,46.9,44.1,29.8,24.0,14.7.HR-ESIMS m/z 419.1498[M–H]–(calcd.for C26H19N4O2,419.1508).
化合物67的制备
按照化合物2的制备方法,以化合物66(10.2mg,23.8μmol)和NaHCO3(4mg,47.6μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=5:1(v/v)洗脱得黄色荧光粉末6-羟甲基-12-乙基-13-氰丙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(67)10.5mg,收率96%。1H NMR(600MHz,CDCl3)δ9.25(d,1H,J=7.7Hz,Ar-H),9.23(d,1H,J=7.7Hz,Ar-H),7.65-7.61(m,4H,Ar-H),7.47(t,1H,J=7.7Hz,Ar-H),7.44(t,1H,J=6.6Hz,Ar-H),5.41(d,2H,J=7.7Hz,N-CH 2-OH),4.84(t,2H,J=7.7Hz,N-CH 2-(CH2)2CN),4.68(q,2H,J=7.7Hz,N-CH 2-CH3),3.17(t,1H,J=7.7Hz,-OH),1.74-1.78(m,4H,NCH2-CH 2-CH 2-CN),1.11(t,3H,J=7.7Hz,CH3).13C NMR(150MHz,CDCl3)δ169.1×2,144.9,144.1,133.7,133.4,132.7,128.1,128.0,126.3,126.0,124.6,124.5,122.6,122.3,122.2,121.6,120.3,118.2,112.2,111.8,61.8,46.9,44.1,24.0,14.7,13.6.HR-ESIMS m/z473.1575[M+H]+(calcd.for C27H22N4O3Na,473.1590).
化合物68的制备
按照化合物64的制备方法,以化合物5(40mg,0.092mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色荧光粉末12-乙基-13-氰丁基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(68)28.7mg,收率72%。1H NMR(600MHz,CDCl3)δ9.26(d,1H,J=7.7Hz,Ar-H),9.24(d,1H,J=7.7Hz,Ar-H),7.72(s,1H,imide-NH),7.64(t,1H,J=7.3Hz,Ar-H),7.63(d,1H,J=7.4Hz,Ar-H),7.62(t,1H,J=7.7Hz,Ar-H),7.60(d,1H,J=7.8Hz,Ar-H),7.463(dt,1H,J=7.6Hz,1.1Hz,Ar-H),7.462(dt,1H,J=7.4Hz,0.9Hz,Ar-H),4.75(t,2H,J=6.9Hz,N-CH 2-(CH2)3CN),4.65(q,2H,J=7.4Hz,N-CH 2-CH3),1.99(t,2H,J=6.8Hz,N(CH2)3-CH 2-CN),1.67(m,2H,NCH2-CH 2-(CH2)2CN),1.10(t,3H,J=7.4Hz,-CH2-CH 3),1.00-0.98(m,2H,N(CH2)2-CH 2-CH2CN).13C NMR(150MHz,CDCl3)δ169.5×2,145.0,144.1,133.8,133.6,127.9,127.8,126.3,126.1,124.7,124.4,122.3,122.2,122.0,121.7,121.3,121.2,118.9,112.1,111.9,47.6,44.1,29.8,27.3,22.3,16.7.HR-ESIMS m/z 433.1663[M–H]–(calcd.for C27H21N4O2,433.1665).
化合物69的制备
按照化合物2的制备方法,以化合物68(9.8mg,22.6μmol)和NaHCO3(4mg,45.2μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得黄色荧光粉末6-羟甲基-12-乙基-13-氰丁基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(70)10mg,收率95%。1H NMR(600MHz,CDCl3)δ9.26(d,1H,J=7.7Hz,Ar-H),9.24(d,1H,J=8.8Hz,Ar-H),7.64-7.60(m,4H,Ar-H),7.45(t,2H,J=7.7Hz,Ar-H),5.41(d,2H,J=7.2Hz,N-CH 2-OH),4.75(t,2H,J=6.6Hz,N-CH 2-(CH2)3CN),4.66(q,2H,J=6.6Hz,N-CH 2-CH3),3.15(t,1H,J=7.2Hz,-OH),1.97(t,2H,J=7.8Hz,N(CH2)3-CH 2-CN),1.68-1.64(m,2H,NCH2-CH 2-(CH2)2CN),1.10(t,3H,J=6.6Hz,-CH2-CH 3),0.98(m,2H,N(CH2)2-CH 2-CH2CN).13C NMR(150MHz,CDCl3)δ169.2×2,144.9,144.1,133.8,133.6,128.9,127.9,127.8,126.2,126.0,124.7,122.6,122.3×2,122.1,120.5,119.3,118.9,112.2,111.9,61.8,47.5,44.1,27.3,22.3,16.7,13.6.ESI-MS m/z 465.3[M+H]+.
化合物70的制备
按照化合物64的制备方法,以化合物7(60mg,0.15mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得黄色荧光粉末12,13-二氰甲基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(70)25mg,收率42%。1H NMR(600MHz,DMSO-d6)δ11.40(s,1H,imide-NH),9.12(d,2H,J=7.8Hz,Ar-H),8.02(d,2H,J=8.3Hz,Ar-H),7.77(dt,2H,J=7.3Hz,1.1Hz,Ar-H),7.56(t,2H,J=7.3Hz,Ar-H),5.75(s,4H,-CH2 -CN).13C NMR(150MHz,DMSO-d6)δ170.0×2,144.5×2,132.3×2,128.6×2,125.1×2,123.6×2,123.2×2,122.2×2,121.5×2,115.3×2,112.8×2,37.4×2.ESI-MS m/z 404.1[M+H]+.
化合物71的制备
按照化合物64的制备方法,以化合物2,3-di(1-cyanoethyl-1H-indol-3-yl)maleimide(30mg,0.07mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得黄色荧光粉末12,13-二氰乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(71)12mg,收率40%。1H NMR(600MHz,DMSO-d6)δ11.27(s,1H,imide-NH),9.14(d,2H,J=8.3Hz,Ar-H),8.00(d,2H,J=8.2Hz,Ar-H),7.69(t,2H,J=7.7Hz,Ar-H),7.49(d,2H,J=7.7Hz,Ar-H),5.07(t,4H,J=6.6Hz,N-CH2 -CH2CN),2.64(4H,t,J=6.6Hz,NCH2-CH2 -CN).13C NMR(150MHz,DMSO-d6)δ171.2×2,143.8×2,133.6×2,128.3×2,125.6×2,124.5×2,122.7×2,121.8×2,121.2×2,118.2×2,113.6×2,44.3×2,21.5×2.ESI-MS m/z430.0[M–H]–.
化合物72的制备
以化合物65的制备方法,以化合物9(45mg,0.09mmol)为原料制备,经硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得黄色荧光粉末12,13-二氰丁基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(74)17mg,收率43%。1H NMR(600MHz,DMSO-d6)δ11.18(s,1H,imide-NH),9.15(d,2H,J=7.8Hz,Ar-H),7.93(d,2H,J=8.2Hz,Ar-H),7.66(dt,2H,J=7.8Hz,1.0Hz,Ar-H),7.44(t,2H,J=7.8Hz,Ar-H),4.78(t,4H,J=7.4Hz,N-CH 2-(CH2)3CN),2.26(t,4H,J=7.3Hz,N(CH2)3-CH2 -CN),1.53-1.50(m,4H,NCH2-CH2 -(CH2)2CN),1.11-1.09(4H,m,N(CH2)2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ171.2×2,144.6×2,133.4×2,128.0×2,125.4×2,123.7×2,122.0×2,121.4×2,120.7×2,120.3×2,113.4×2,48.1×2,27.6×2,22.5×2,16.1×2.ESI-MS m/z 486.1[M–H]–.
化合物73的制备
i)12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(73a)的制备
将化合物24d(400mg,1.13mmol)、DDQ(282mg,1.24mmol)和p-TsOH(214mg,1.13mmol)以100mL苯溶解,N2保护条件下回流30min,蒸干溶剂,100mL乙酸乙酯重新溶解,分别用饱和NaHSO3溶液、水、盐洗,有机层用无水Na2SO4干燥,蒸干后硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得黄色粉末(73a)280mg,收率70%。1H NMR(500MHz,DMSO-d6)δ11.96(s,1H,indole-NH),10.99(s,1H,imido-NH),9.12(d,1H,J=8.1Hz,Ar-H),9.07(d,1H,J=8.1Hz,Ar-H),7.79(d,2H,J=8.0Hz,Ar-H),7.59(t,1H,J=7.9Hz,Ar-H),7.56(t,1H,J=9.0Hz,Ar-H),7.36(d,1H,J=8.2Hz,Ar-H),7.34(d,1H,J=8.1Hz,Ar-H),4.94(q,2H,J=7.1Hz,-CH2 -CH3),1.42(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.6×2,141.5,141.0,129.6,128.6,127.3,125.1,124.7,121.7,121.4,120.7×2,120.4,120.2,117.2,116.3,112.5,110.2,110.1,39.8,16.1.HR-ESIMS m/z 354.1249[M+H]+(calcd.for C22H16N3O2,354.1243).
ii)6-(2-氨乙基)-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(73)的制备
用100mL 10%KOH水溶液悬浮化合物73a(470mg,1.33mmol),110℃回流1.5h,溶解为淡黄色澄清溶液,冷却至室温,2N盐酸酸化,乙酸乙酯萃取(100mL×3次),将有机层蒸干得465mg粗品12-乙基-12,13-二氢呋喃[3,4-c]吲哚[2,3-a]咔唑-5,7-二酮(73b),该化合物溶解性极差,但反应完全、产物单一,所以未经分离直接投入下步反应。按照化合物14的制备方法,以所得粗品73b和2mL乙二胺为原料,制得黄色粉末(73)510mg,收率97%。1H NMR(500MHz,DMSO-d6)δ9.03(t,2H,J=8.5Hz,Ar-H),7.80(d,1H,J=8.1Hz,Ar-H),7.74(d,1H,J=8.1Hz,Ar-H),7.57(t,1H,J=7.8Hz,Ar-H),7.53(t,1H,J=7.6Hz,Ar-H),7.33(d,1H,J=8.1Hz,Ar-H),7.31(d,1H,J=8.0Hz,Ar-H),4.86(q,2H,J=6.9Hz,-CH2 -CH3),3.75(t,2H,J=6.1Hz,-NCH2 CH2NH2),2.97(t,2H,J=6.1Hz,-NCH2CH2 -NH2),1.36(t,3H,J=6.9Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ170.1×2,141.5,140.9,129.4,128.3,127.3,127.2,124.9,124.5,121.5,121.2,120.6×2,119.2,119.0,117.2,116.3,112.6,110.1,39.9,39.6,38.7,16.1.HR-ESIMS m/z 397.1671[M+H]+(calcd.for C24H21N4O2,397.1665).
化合物74的制备
按照化合物16的制备方法,以化合物73(510mg,1.26mmol)为原料制得黄色粉末6-(2-氨乙基)-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮盐酸盐(74)540mg,收率99%。1H NMR(600MHz,DMSO-d6)δ12.07(s,1H,indole-NH),9.11(d,1H,J=8.2Hz,Ar-H),9.08(d,1H,J=8.2Hz,Ar-H),7.90(brs,3H,-NH3 +),7.84(d,2H,J=8.1Hz,Ar-H),7.63(t,1H,J=7.1Hz,Ar-H),7.58(t,1H,J=7.7Hz,Ar-H),7.38(t,1H,J=7.8Hz,Ar-H),7.35(t,1H,J=7.7Hz,Ar-H),4.98(q,2H,J=6.9Hz,-CH2 -CH3),3.98(t,2H,J=5.9Hz,N-CH2 -CH2NH3 +),3.19(t,2H,J=6.0Hz,NCH2-CH2 -NH3 +),1.41(t,3H,J=6.9Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ170.1×2,141.6,141.1,129.6,128.5,127.5×2,124.9,124.5,121.6,121.2,120.8,120.8,119.5,119.3,117.3,116.5,112.7,110.4,39.6,38.5,35.8,16.1.HR-ESIMS m/z 397.1670[M-Cl]+(calcd.for C24H21N4O2,397.1665).
化合物75的制备
0℃下,以15mL DMF悬浮NaH于250mL两口烧瓶中,滴加1mL溶解的化合物73a(40mg,0.113mmol),低温反应10min,升至室温反应30min。降温至0℃,滴加ClCH2CH2OH(0.092mL,1.36mol)。55℃油浴下,Ar气保护,冷凝水回流5h,TLC检测反应未完全进行,升温至85℃,反应6h。降至室温,低温下加入10mL甲醇,20mL饱和氯化铵水溶液。乙酸乙酯萃取,有机层以无水Na2SO4干燥,真空蒸干。THF溶解,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得黄色固体6-(2-羟乙基)-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(75)11mg,收率25%。1H NMR(500MHz,DMSO-d6)δ11.95(s,1H,indole-NH),9.09(d,1H,J=8.1Hz,Ar-H),9.08(d,1H,J=8.1Hz,Ar-H),7.79(d,1H,J=7.8Hz,Ar-H),7.77(d,1H,J=7.6Hz,Ar-H),7.59(t,1H,J=7.4Hz,Ar-H),7.56(t,1H,J=7.4Hz,Ar-H),7.35(t,1H,J=8.1Hz,Ar-H),7.33(t,1H,J=7.4Hz,Ar-H),4.90(q,2H,J=6.9Hz,-CH2 -CH3),3.73-3.71(m,2H,-NCH2CH2 -OH),3.69(t,2H,J=5.4Hz,-NCH2 CH2OH),1.41(t,3H,J=6.9Hz,-CH2-CH3 ).13CNMR(125MHz,DMSO-d6)δ170.1×2,141.8,141.0,129.5,128.4,127.3,125.0,124.6,121.6,121.3,120.7×2,119.3,119.1,117.2,116.4,112.5,110.1×2,58.8,40.5,39.5,16.1.HR-ESIMS m/z398.1508[M+H]+(calcd.for C24H20N3O3,398.1505).
化合物76的制备
按照化合物24的制备方法,以化合物73b(49mg,0.14mmol)、4-羟基苄胺(51mg,0.42mmol)和催化量Et3N为原料,制得黄色固体6-(4-羟基苄基)-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(76)15mg,收率23%。1H NMR(600MHz,DMSO-d6)δ9.03(d,1H,J=7.0Hz,Ar-H),9.03(d,1H,J=6.8Hz,Ar-H),7.77(d,1H,J=8.2Hz,Ar-H),7.70(d,1H,J=8.4Hz,Ar-H),7.56(d,1H,J=7.0Hz,Ar-H),7.55(d,1H,J=6.9Hz,Ar-H),7.34(dd,1H,J=7.0Hz,0.9Hz,Ar-H),7.32(dd,1H,J=7.0Hz,0.9Hz,Ar-H),7.20(d,2H,J=8.2Hz,Ar-H),6.71(d,2H,J=8.2Hz,Ar-H),4.81(q,2H,J=7.2Hz,-CH2 -CH3),4.63(s,2H,-NCH2-Ar),1.38(t,3H,J=7.2Hz,-CH2-CH3 ).13CNMR(150MHz,DMSO-d6)δ169.8,169.7,157.2,141.7,141.0,129.6×2,128.5,128.4,127.4,125.0,124.9,124.7,121.5,121.3,120.9,120.8,119.0,118.8,117.4,116.4,115.8×2,112.6,110.1,100.0,40.7,39.6,16.3.ESI-MS m/z 460.1[M+H]+.
化合物77的制备
按照化合物24的制备方法,以化合物73b(35mg,0.098mmol)、4-(2-氨乙基)吗啉(104μL,0.79mmol)和催化量Et3N为原料,制得黄色固体6-(2-(4-吗啉)乙基)-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(77)33mg,收率72%。1H NMR(500MHz,DMSO-d6)δ12.01(s,1H,indole-NH),9.09(t,1H,J=8.1Hz,Ar-H),9.07(t,1H,J=7.9Hz,Ar-H),7.81(d,2H,J=8.0Hz,Ar-H),7.59(t,1H,J=8.0Hz,Ar-H),7.56(t,1H,J=7.8Hz,Ar-H),7.38(d,1H,J=8.0Hz,Ar-H),7.35(t,1H,J=8.0Hz,Ar-H),4.94(q,2H,J=7.1Hz,-CH2 -CH3),3.79(t,2H,J=6.2Hz,imide-NCH2 CH2-),3.50(t,4H,J=4.5Hz,morpholine-N(CH2-CH 2)2O),2.62(t,2H,J=6.2Hz,imide-NCH2CH2 -),2.46(t,4H,J=4.5Hz,morpholine-N(CH 2-CH2)2O),1.37(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ170.0×2,141.7,140.9,129.4,128.5,127.5,127.3,125.0,124.7,121.6,121.5,121.4,121.2,120.9,119.0,117.4,116.5,112.7,110.2,66.8×2,56.6,53.7×2,39.6,34.9,16.2.HR-ESIMS m/z 467.2088[M+H]+(calcd.for C28H27N4O3,467.2083).
化合物78的制备
按照化合物24的制备方法,以化合物73b(100mg,0.282mmol)、N,N-二甲基乙二胺(247.7μL,2.256mmol)和催化量Et3N为原料,制得黄色固体6-(2-(N,N-二甲氨基乙基))-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(78)95.7mg,收率80%。1H NMR(600MHz,DMSO-d6)δ11.88(s,1H,indole-NH),9.01(d,2H,J=7.4Hz,Ar-H),7.76(d,1H,J=8.1Hz,Ar-H),7.69(d,1H,J=8.3Hz,Ar-H),7.56(d,1H,J=8.1Hz,Ar-H),7.53(d,1H,J=7.9Hz,Ar-H),7.33(td,2H,J=7.8Hz,2.3Hz,Ar-H),4.80(q,2H,J=7.1Hz,-CH2 -CH3),3.63(t,2H,J=6.4Hz,imide-NCH2 CH2N(CH3)2),2.49(t,2H,J=6.4Hz,imide-NCH2CH2 -N(CH3)2),2.18(s,6H,-N(CH3)2),1.38(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ169.9×2,141.6,141.0,129.5,128.4,127.4×2,125.0,124.8×2,121.6,121.3,120.7,119.1,118.9,117.3,116.4,112.6,110.1,57.5,45.7×2,39.7,35.7,16.2.HR-ESIMS m/z 425.1988[M+H]+(calcd.for C26H25N4O2,425.1978).
化合物79的制备
按照化合物24的制备方法,以化合物73b(80mg,0.226mmol)、2-(2-氨乙基)吡啶(135.3μL,1.13mmol)和催化量Et3N为原料,制得黄色固体6-(2-(2-吡啶)乙基)-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(79)25mg,收率64%。72.5mg,收率70%。1H NMR(500MHz,DMSO-d6)δ11.92(s,1H,indole-NH),9.02(d,1H,J=6.5Hz,Ar-H),9.01(t,1H,J=6.5Hz,Ar-H),8.43(d,1H,J=3.2Hz,Ar-H),7.77(t,2H,J=7.4Hz),7.66(t,1H,J=8.0Hz,Ar-H),7.58(t,1H,J=8.0Hz,Ar-H),7.55(t,1H,J=7.8Hz,Ar-H),7.33-7.28(m,3H,Ar-H),7.18(t,1H,J=6.3Hz,Ar-H),4.88(q,2H,J=7.0Hz,-CH2 -CH3),4.00(t,2H,J=6.5Hz,imide-NCH2 CH2-),3.13(t,2H,J=6.5Hz,imide-NCH2CH2 -),1.39(t,3H,J=7.0Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ169.8×2,159.0,149.7,141.6,141.0,137.1,129.5,128.4,127.3,125.0,124.7,123.8,123.7,122.9,122.2,121.6,121.3,120.7,119.1,118.9,117.3,116.4,112.5,110.1,39.6,37.7,36.9,16.2.HR-ESIMSm/z 459.1831[M+H]+(calcd.for C29H23N4O2,459.1821).
化合物80的制备
按照化合物64的制备方法,以化合物62(41mg,0.09mmol)为原料制得黄色固体2-异戊烯基-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(80)38mg,收率90%。1H NMR(600MHz,DMSO-d6)δ11.80(s,1H,indole-NH),10.95(s,1H,imide-NH),9.06(d,1H,J=7.6Hz,Ar-H),8.88(d,1H,J=8.1Hz,Ar-H),7.75(d,1H,J=8.3Hz,Ar-H),7.58(td,1H,J=7.1Hz,1.1Hz,Ar-H),7.50(s,1H,Ar-H),7.33(t,1H,J=7.5Hz,Ar-H),7.13(dd,1H,J=8.1Hz,1.3Hz,Ar-H),5.40(t,1H,J=7.5Hz,ArCH2CH=C(CH3)2),4.87(q,2H,J=7.1Hz,-CH2 -CH3),3.48(d,2H,J=7.5Hz,ArCH2 CH=C(CH3)2),1.75(s,6H,ArCH2CH=C(CH3 )2),1.39(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ170.7,170.6,141.0,140.0,139.9,131.5,128.6,127.5,126.2,124.0,123.5,122.8,120.7,119.6,119.1,118.8,118.4,116.3,115.1,110.3,109.0,106.5,33.6,28.3,25.1,17.2,15.2.HR-ESIMSm/z 422.1879[M+H]+(calcd.for C27H24N3O2,422.1869).
化合物81的制备
按照化合物2的制备方法,以化合物80(30mg,0.071mmol)和甲醛溶液(3mL,质量分数37%)为原料制得黄色固体2-异戊烯基-6-羟甲基-12-乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(81)13mg,收率25%。1H NMR(600MHz,DMSO-d6)δ11.78(s,1H,indole-NH),9.03(d,1H,J=7.8Hz,Ar-H),8.86(d,1H,J=8.1Hz,Ar-H),7.72(d,1H,J=8.3Hz,Ar-H),7.57(t,1H,J=7.0Hz,Ar-H),7.51(s,1H,Ar-H),7.34(t,1H,J=7.4Hz,Ar-H),7.14(d,1H,J=8.2Hz,Ar-H),6.28(t,1H,J=7.0Hz,-CH2OH),5.44(t,1H,J=7.6Hz,ArCH2CH=C(CH3)2),4.92(d,2H,J=6.1Hz,-CH2 OH),4.80(q,2H,J=7.2Hz,-CH2 -CH3),3.53(d,2H,J=7.5Hz,ArCH2 CH=C(CH3)2),1.78(s,6H,ArCH2CH=C(CH3 )2),1.39(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ168.6×2,141.3,140.3,140.2,131.8,128.8,127.7,126.6,124.1,123.6,123.0×2,120.8,112.0,118.6,118.1,117.7,116.5,115.4,110.6,109.3,59.4,38.8,33.9,25.4,17.5,15.4.HR-ESIMS m/z 450.1821[M-H]-(calcd.for C28H24N3O3,450.1812).
化合物82的制备
i)N-甲基-2,3-二溴马来酰亚胺(82a)的制备
在50mL两口瓶中,将NaH(30mg,0.75mmol,质量分数60%,分散于石蜡中)用5mLDMF悬浮搅拌,–5℃下滴加5mL DMF溶解的2,3-二溴马来酰亚胺(127.5mg,0.5mmol),低温反应30min后,滴加碘甲烷(47μL,0.75mmol),低温反应30min,滴加饱和NH4Cl溶液终止反应,CH2Cl2萃取,有机层蒸干,硅胶柱色谱分离、石油醚:乙酸乙酯=30:1(v/v)洗脱得白色晶体(82a)92mg,收率69%。1H NMR(600MHz,CDCl3)δ3.12(s,3H,-CH3).13C NMR(150MHz,CDCl3)δ164.1×2,129.5×2,25.6.ESI-MS m/z 267.9[M+H]+.
ii)N-甲基-2,3-二(6-氯-3-吲哚)马来酰亚胺(82)的制备
在50mL两口瓶中放置镁丝(200mg,8.35mmol),室温下用5mLTHF悬浮搅拌,滴加溴代乙烷(620μL,8.35mmol),室温反应20min,升至45℃继续反应30min,滴加8mL甲苯溶解的6-氯吲哚(1.27g,8.35mmol),反应1h,缓慢滴加8mL甲苯溶解的82a(448mg,1.67mmol),滴毕升至110℃回流2h,降至–5℃下,滴加饱和NH4Cl溶液终止反应,乙酸乙酯萃取,有机层浓缩,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色固体粉末(82)300mg,收率44%。1HNMR(500MHz,DMSO-d6)δ11.80(s,2H,indole-NH),7.80(s,2H,Ar-H),7.43(s,2H,Ar-H),6.71(d,2H,J=8.6Hz,Ar-H),6.65(d,2H,J=8.6Hz,Ar-H),3.02(s,3H,-CH3).13C NMR(125MHz,DMSO-d6)δ171.9×2,136.8×2,130.7×2,127.4×2,126.8×2,124.4×2,122.3×2,120.1×2,112.0×2,106.0×2,24.4.HR-ESIMS m/z 410.0467[M+H]+(calcd.forC21H14N3O2Cl2,410.0463).
化合物83的制备
在25mL两口瓶中,用10mL DMF溶解化合物82(257mg,0.63mmol),-5℃搅拌条件下加入NaH(28mg,0.69mmol,质量分数60%,分散于石蜡中),低温反应30min后,缓慢滴加EtI(108mg,0.69mmol),低温反应30min。滴加饱和NH4Cl溶液终止反应,乙酸乙酯萃取,有机层浓缩,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得橙红色粉末N-甲基-2-(1-乙基-6-氯-3-吲哚)-3-(6-氯-3-吲哚)马来酰亚胺(83)100mg,收率38%。1H NMR(500MHz,DMSO-d6)δ11.84(d,1H,J=2.2Hz,indole-NH),7.83(d,1H,J=2.2Hz,Ar-H),7.79(s,1H,Ar-H),7.64(d,1H,J=1.6Hz,Ar-H),7.45(d,1H,J=1.7Hz,Ar-H),6.81(d,1H,J=8.8Hz,Ar-H),6.72(dd,1H,J=8.3Hz,1.7Hz,Ar-H),6.66-6.64(m,2H,Ar-H),4.25(q,2H,J=7.1Hz,-CH 2-CH3),3.03(s,3H,N-CH3),1.31(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.0,171.9,137.1,136.5,132.9,131.0,127.7,127.3,127.0,126.9,125.1,124.3,122.9,122.6,120.5,120.3,112.2,110.8,106.0,105.6,41.4,24.6,15.8.HR-ESIMSm/z 438.0780[M+H]+(calcd.for C23H18N3O2Cl2,438.0776).
化合物84的制备
i)2-(1-乙基-6-氯-3-吲哚)-3-(6-氯-3-吲哚)马来酸酐(84a)的制备
在50mL单口瓶中,用10mL乙醇悬浮化合物83(100mg,0.23mmol),加入10mL 5M的KOH溶液,78℃下回流8h后冷却至室温,滴加6N盐酸酸化,乙酸乙酯萃取,有机层用无水硫酸钠干燥,真空浓缩,硅胶柱色谱分离、二氯甲烷洗脱得橙红色固体(84a)58mg,收率60%。1HNMR(600MHz,DMSO-d6)δ12.06(d,1H,J=2.8Hz,indole-NH),7.92(d,1H,J=3.3Hz,Ar-H),7.88(s,1H,Ar-H),7.69(d,1H,J=1.6Hz,Ar-H),7.49(d,1H,J=1.6Hz,Ar-H),6.85(d,1H,J=8.8Hz,Ar-H),6.79(dd,1H,J=8.2Hz,1.6Hz,Ar-H),6.73(dd,1H,J=8.2Hz,1.6Hz,Ar-H),6.70(d,1H,J=8.8Hz,Ar-H),4.26(q,2H,J=7.1Hz,-CH2 -CH3),1.31(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.8,166.7,137.2,136.7,134.0,132.3,128.8,128.1,127.7,127.4,124.7,124.0,123.2,122.9,121.0,120.7,112.5,111.1,105.4,104.8,41.6,15.7.ESI-MS m/z 425.0/427.0[M+H]+.
ii)2-(1-乙基-6-氯-3-吲哚)-3-(6-氯-3-吲哚)马来酰亚胺(84)的制备
按照化合物24c的制备方法,以化合物84a(53mg,0.125mmol)、HMDS(2.6mL,12.5mmol)和MeOH(0.25mL,6.25mmol)为原料,制得橙红色粉末(84)52mg,收率98%。1H NMR(500MHz,DMSO-d6)δ11.80(s,1H,indole-NH),10.97(s,1H,imide-NH),7.81(d,1H,J=2.8Hz,Ar-H),7.79(s,1H,Ar-H),7.62(s,1H,Ar-H),7.44(s,1H,Ar-H),6.80(d,1H,J=8.8Hz,Ar-H),6.71(dd,1H,J=8.8Hz,1.1Hz,Ar-H),6.69(d,1H,J=8.8Hz,Ar-H),6.65(dd,1H,J=8.8Hz,1.7Hz,Ar-H),4.25(q,2H,J=7.1Hz,-CH2 -CH3),1.31(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ173.2×2,137.0,136.5,132.9,130.9,128.3,127.7,127.2,126.9,125.2,124.5,122.9,122.6,120.4,120.2,112.1,110.0,106.0,105.6,41.4,15.8.HR-ESIMS m/z 424.0629[M+H]+(calcd.for C22H16N3O2Cl2,424.0620).
化合物85的制备
按照化合物2的制备方法,以化合物84(20mg,47μmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(11.9mg,140μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-6-氯-3-吲哚)-3-(1-羟甲基-6-氯-3-吲哚)马来酰亚胺(85)22mg,收率97%。1H NMR(500MHz,DMSO-d6)δ8.00(s,1H,Ar-H),7.80(s,1H,Ar-H),7.68(s,1H,Ar-H),7.64(s,1H,Ar-H),6.86(d,1H,J=8.6Hz,Ar-H),6.74(t,1H,J=7.2Hz,indole-CH2-OH),6.70(d,1H,J=8.7Hz,Ar-H),6.67(d,1H,J=8.6Hz,Ar-H),6.55(d,1H,J=8.6Hz,Ar-H),6.33(t,1H,J=7.0Hz,imide-CH2-OH),5.60(d,2H,J=7.2Hz,indole-CH2 -OH),4.95(d,2H,J=7.0Hz,imide-CH2 -OH),4.25(q,2H,J=7.2Hz,-CH2 -CH3),1.29(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.3×2,136.6×2,133.5,133.1,127.8,127.4×2,127.3,125.3,125.2,122.9,122.7,120.8,120.6,111.6,110.9,105.7,105.4,69.9,60.9,41.5,15.8.HR-ESIMS m/z 506.0656[M+Na]+(calcd.forC24H19N3O4Cl2Na,506.0650).
化合物86的制备
按照化合物82的制备方法,以化合物82a(710mg,2.64mmol)、Mg(317mg,13.2mmol)、溴代乙烷(982μL,13.2mmol)和4-溴吲哚(2g,132mmol)为原料制备,硅胶柱色谱分离、二氯甲烷:乙酸乙酯=9:1(v/v)洗脱得固体N-甲基-2,3-二(4-溴-3-吲哚)马来酰亚胺(86)500mg,收率30%。1H NMR(500MHz,DMSO-d6)δ11.84(s,2H,indole-NH),7.84(d,2H,J=7.4Hz,Ar-H),7.42(d,2H,J=8.1Hz,Ar-H),7.18(d,2H,J=7.5Hz,Ar-H),7.02(t,2H,J=7.8Hz,Ar-H),3.07(s,3H,-CH3).13C NMR(125MHz,DMSO-d6)δ172.3×2,137.8×2,137.3×2,130.4×2,127.1×2,126.1×2,123.6×2,113.9×2,112.4×2,104.8×2,24.8.ESI-MSm/z 519.9/521.9/523.9[M+Na]+.
化合物87的制备
按照化合物83的制备方法,以化合物86(506mg,1.02mmol)、NaH(81mg,2.04mmol,质量分数60%,分散于石蜡中)和碘乙烷(90μL,1.2mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得到橙红色固体N-甲基-2-(1-乙基-4-溴-3-吲哚)-3-(4-溴-3-吲哚)马来酰亚胺(87)182mg,收率34%。1H NMR(500MHz,DMSO-d6)δ11.85(s,1H,indole-NH),7.92(s,1H,Ar-H),7.85(d,1H,J=2.7Hz,Ar-H),7.52(d,1H,J=8.3Hz,Ar-H),7.41(d,1H,J=8.1Hz,Ar-H),7.19(dd,2H,J=7.2Hz,,1.6Hz,Ar-H),7.06(t,1H,J=7.9Hz,Ar-H),7.02(t,1H,J=7.8Hz,Ar-H),4.22(q,2H,J=7.2Hz,-CH2 -CH3),3.06(s,3H,-NCH3),1.34(t,J=7.2Hz,3H,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.3×2,137.7×2,135.3,134.7,132.7,130.4,126.3,126.1,124.2,124.1,123.6,123.5,114.1,113.9,111.9,110.4,104.2,103.5,41.4,24.8,15.8.ESI-MS m/z 525.9/527.9/529.9[M+H]+.
化合物88的制备
i)2-(1-乙基-4-溴-3-吲哚)-3-(4-溴-3-吲哚)马来酸酐(88a)的制备
按照化合物84a的制备方法,以化合物87(100mg,0.23mmol)为原料制备,硅胶柱色谱(二氯甲烷洗脱)得橙红色固体(88a)58mg,收率60%。1H NMR(600MHz,DMSO-d6)δ12.06(s,1H,indole-NH),8.00(s,1H,Ar-H),7.95(d,1H,J=2.7Hz,Ar-H),7.57(d,1H,J=8.2Hz,Ar-H),7.46(d,1H,J=8.1Hz,Ar-H),7.25(dd,2H,J=7.2Hz,7.1Hz,Ar-H),7.10(t,1H,J=7.9Hz,Ar-H),7.06(t,1H,J=7.9Hz,Ar-H),4.25(q,2H,J=7.2Hz,-CH2 -CH3),1.35(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.7×2,137.8,137.3,136.9,136.3,133.6,131.4,125.8,125.6,124.7,124.6,124.0×2,113.9,113.7,112.3,110.7,102.9,102.2,41.6,15.8.ESI-MS m/z 513.1/515.0/517.1[M+H]+.
ii)2-(1-乙基-4-溴-3-吲哚)-3-(4-溴-3-吲哚)马来酰亚胺(88)的制备
按照化合物24c的制备方法,以化合物88a(126mg,0.32mmol)、HMDS(6.7mL,32mmol)和MeOH(0.64mL,16mmol)为原料制备,硅胶柱色谱分离、二氯甲烷洗脱得橙红色粉末88(118mg,收率94%)。1H NMR(500MHz,DMSO-d6)δ11.83(s,1H,indole-NH),10.98(s,1H,imide-NH),7.92(s,1H,Ar-H),7.85(d,1H,J=2.7Hz,Ar-H),7.52(t,1H,J=8.0Hz,Ar-H),7.41(d,1H,J=8.0Hz,Ar-H),7.20(t,1H,J=7.5Hz,Ar-H),7.18(d,1H,J=7.5Hz,Ar-H),7.06(d,1H,J=7.9Hz,Ar-H),7.01(t,1H,J=7.8Hz,Ar-H),4.21(q,2H,J=7.2Hz,-CH2 -CH3),1.35(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ173.5×2,137.7,137.2,135.8,135.3,132.6,130.3,126.4,124.1,124.0,123.5,114.2,113.9,113.7,111.9,110.3,104.4,103.7,100.0,41.4,15.8.ESI-MS m/z 512.1/514.0/516.1[M+H]+.
化合物89的制备
按照化合物1的制备方法,以化合物88(36mg,70.5μmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(30mg,352μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得橙黄色固体N-羟甲基-2-(1-乙基-4-溴-3-吲哚)-3-(1-羟甲基-4-溴-3-吲哚)马来酰亚胺(89)39mg,收率97%。1H NMR(500MHz,DMSO-d6)δ7.99(s,1H,Ar-H),7.92(s,1H,Ar-H),7.61(d,1H,J=8.2Hz,Ar-H),7.53(d,1H,J=8.2Hz,Ar-H),7.24(d,1H,J=7.8Hz,Ar-H),7.21(d,1H,J=7.6Hz,Ar-H),7.09(t,1H,J=8.0Hz,Ar-H),7.06(t,1H,J=7.6Hz,Ar-H),6.69(t,1H,J=7.3Hz,-CH2OH),6.40(t,1H,J=7.0Hz,-CH2OH),5.56(d,2H,J=7.3Hz,-CH2 OH),4.97(d,2H,J=7.0Hz,-CH2 OH),4.02(q,2H,J=7.1Hz,-CH2 -CH3),1.17(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.6×2,137.2,137.1,135.5,135.0,133.2,132.9,126.8,126.3,124.7,124.2,123.8,123.6,114.0,113.9,111.0,110.4,103.9,103.3,69.7,60.3,41.5,15.8.ESI-MS m/z 554.1/556/1/558.1[M–H2O+H]+.
化合物90的制备
按照化合物82的制备方法,以化合物5-溴吲哚(2g,13.2mmol)、Mg(317mg,13.2mmol)和化合物82a(710mg,2.64mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得橙红色固体N-甲基-2,3-二(5-溴-3-吲哚)马来酰亚胺(90)500mg,收率30%。1H NMR(500MHz,DMSO-d6)δ11.91(s,2H,indole-NH),7.80(s,2H,Ar-H),7.36(d,2H,J=8.6Hz,Ar-H),7.10(d,2H,J=8.6Hz,Ar-H),6.84(s,2H,Ar-H),3.04(s,3H,-CH3).13C NMR(125MHz,DMSO-d6)δ171.9×2,135.2×2,130.9×2,127.5×2,127.3×2,124.7×2,123.6×2,114.2×2,112.4×2,105.5×2,24.5.HR-ESIMS m/z 497.9458[M+H]+(calcd.forC21H14N3O2Br2,497.9453).
化合物91的制备
按照化合物84的制备方法,以化合物90(506mg,1.2mmol)、NaH(81mg,2.04mmol,质量分数60%,分散于石蜡中)和EtI(90μL,1.2mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得橙红色固体N-甲基-2-(1-乙基-5-溴-3-吲哚)-3-(5-溴-3-吲哚)马来酰亚胺(91)200mg,收率36%。1H NMR(500MHz,DMSO-d6)δ11.95(s,1H,indole-NH),7.88(s,1H,Ar-H),7.75(d,1H,J=1.4Hz,Ar-H),7.48(d,1H,J=8.1Hz,Ar-H),7.37(d,1H,J=8.5Hz,Ar-H),7.18(d,1H,J=8.5Hz,Ar-H),7.09(d,1H,J=7.9Hz,Ar-H),6.99(s,1H,Ar-H),6.68(s,1H,Ar-H),4.24(q,2H,J=7.1Hz,-CH2 -CH3),3.03(s,3H,-CH3),1.31(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.0,171.9,135.4,134.8,133.0,131.3,128.1,127.9,127.1,126.8,124.8×2,124.0,123.7,114.3,112.8×2,112.6,105.6,105.0,41.5,24.6,16.0.HR-ESIMS m/z 525.9776[M+H]+(calcd.for C23H18N3O2Br2,525.9766).
化合物92的制备
i)2-(1-乙基-5-溴-3-吲哚)-3-(5-溴-3-吲哚)马来酸酐(92a)的制备
按照化合物84a的制备方法,以化合物91(120mg,0.229mmol)为原料制备,硅胶柱色谱分离、二氯甲烷洗脱得红色固体(92a)88mg,收率75%。1H NMR(600MHz,DMSO-d6)δ12.16(s,1H,indole-NH),7.97(d,1H,J=1.4Hz,Ar-H),7.87(s,1H,Ar-H),7.53(d,1H,J=8.7Hz,Ar-H),7.40(d,1H,J=8.5Hz,Ar-H),7.23(d,1H,J=8.8Hz,Ar-H),7.16(d,1H,J=8.4Hz,Ar-H),7.01(s,1H,Ar-H),6.72(s,1H,Ar-H),4.28(q,2H,J=7.2Hz,-CH2 -CH3),1.33(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.8,166.7,135.5,134.9,134.0,132.4,129.1,128.0,127.7,126.8,125.2×2,124.2,124.0,114.6,113.3,113.0×2,104.8,104.2,41.7,16.0.ESI-MS m/z ESI-MS m/z 512.9/514.9/516.9[M+H]+.
ii)2-(1-乙基-5-溴-3-吲哚)-3-(5-溴-3-吲哚)马来酰亚胺(92)的制备
按照化合物23c的制备方法,以化合物92a(88mg,0.172mmol)、HMDS(4mL,17.2mmol)和MeOH(0.5mL,8.6mmol)为原料制备,硅胶柱色谱分离、二氯甲烷洗脱得红色固体92(85mg,收率97%)。1H NMR(500MHz,DMSO-d6)δ11.93(s,1H,indole-NH),10.98(s,1H,imide-NH),7.86(d,1H,J=2.8Hz,Ar-H),7.77(s,1H),7.48(d,1H,J=8.7Hz,Ar-H),7.35(d,1H,J=8.5Hz,Ar-H),7.15(dd,1H,J=8.7Hz,1.8Hz,Ar-H),7.08(dd,1H,J=8.6Hz,1.9Hz,Ar-H),6.94(d,1H,J=7.8Hz,Ar-H),6.68(d,1H,J=1.7Hz,Ar-H),4.25(q,2H,J=7.2Hz,-CH2 -CH3),1.32(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ173.2,173.1,135.3,134.7,133.0,131.2,128.5,128.1,127.4,127.2,124.8,124.7,123.9,123.8,114.3,112.8,112.7,112.5,105.4,105.0,41.5,16.0.HR-ESIMS m/z 511.9617[M+H]+(calcd.for C22H16N3O2Br2,511.9609).
化合物93的制备
按照化合物2的制备方法,以化合物92(55mg,107.6μmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(45mg,538μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得橙黄色固体N-羟甲基-2-(1-乙基-5-溴-3-吲哚)-3-(1-羟甲基-5-溴-3-吲哚)马来酰亚胺(93)58mg,收率95%。1H NMR(500MHz,DMSO-d6)δ8.04(s,1H,Ar-H),7.71(s,1H,Ar-H),7.55(d,1H,J=8.7Hz,Ar-H),7.50(d,1H,J=8.7Hz,Ar-H),7.18(dt,2H,J=8.9Hz,1.5Hz,Ar-H),7.12(d,1H,Ar-H,J=1.5Hz),6.58(d,1H,Ar-H J=1.5Hz),5.60(s,2H,indole-CH2 -OH),4.97(s,2H,imide-CH2 -OH),4.23(q,2H,J=7.1Hz,-CH2 -CH3),3.78(s,1H,N-CH2-OH),3.15(s,1H,N-CH2-OH),1.29(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.2×2,134.9,134.8,133.9,133.2,128.2,128.1,127.6,127.5,127.0,125.0,124.1,124.0,113.5,113.2,113.1,112.8,104.9,104.8,69.9,60.9,41.5,16.0.HR-ESIMSm/z 593.9647[M+Na]+(calcd.for C24H19N3O4Br2Na,593.9640).
化合物94的制备
按照化合物82的制备方法,以6-溴吲哚(850mg,5.58mmol)、Mg(134mg,5,58mmol)和化合物82(300mg,1.12mmol)为原料,制得橙红色固体N-甲基-2,3-二(6-溴-3-吲哚)马来酰亚胺(94)180mg,收率30%。1H NMR(600MHz,DMSO-d6)δ11.83(s,2H,indole-NH),7.78(d,2H,J=7.7Hz,Ar-H),7.58(d,2H,J=7.7Hz,Ar-H),6.78(d,1H,J=7.8Hz,Ar-H),6.77(d,1H,J=7.8Hz,Ar-H),6.68(s,1H,Ar-H),6.67(s,1H,Ar-H),3.02(s,3H,-CH3).13C NMR(150MHz,DMSO-d6)δ172.0×2,137.4×2,130.8×2,127.5×2,124.8×2,122.9×2,122.8×2,115.1×2,115.0×2,106.1×2,24.6.HR-ESIMS m/z 497.9458[M+H]+(calcd.forC21H14N3O2Br2,497.9453).
化合物95的制备
按照化合物83的制备方法,以化合物94(155mg,0.312mmol)、NaH(14mg,0.343mmol,质量分数60%,分散于石蜡中)和EtI(28μL,0.343mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得橙红色固体N-甲基-2-(1-乙基-6-溴-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(95)56mg,收率34%。1H NMR(500MHz,DMSO-d6)δ11.81(s,1H,indole-NH),7.79(s,1H,Ar-H),7.76(s,1H,Ar-H),7.57(s,1H,Ar-H),6.82(d,1H,J=8.6Hz,Ar-H),6.75(t,2H,J=7.7Hz,Ar-H),6.61(d,1H,J=8.6Hz,Ar-H),4.24(q,2H,J=7.2Hz,-CH2 -CH3),3.02(s,3H,-NCH3),1.29(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.9,171.8,137.4,136.8,132.6,130.8,127.6,126.9,125.2,124.4,123.1,122.9,122.8,122.7,115.2,115.0,114.9,113.5,105.9,105.4,41.3,24.4,15.7.HR-ESIMSm/z 525.9771[M+H]+(calcd.for C23H18N3O2Br2,525.9766).
化合物96的制备
i)2-(1-乙基-6-溴-3-吲哚)-3-(6-溴-3-吲哚)马来酸酐(96a)的制备
按照化合物84a的制备方法,以化合物95(240mg,0.46mmol)为原料制备,硅胶柱色谱分离、二氯甲烷洗脱得橙红色固体96a(144mg,收率61%)。1H NMR(600MHz,DMSO-d6)δ12.04(s,1H,indole-NH),7.89(d,1H,J=1.5Hz,Ar-H),7.87(s,1H,Ar-H),7.82(s,1H,Ar-H),7.63(s,1H,Ar-H),6.91(d,1H,J=8.6Hz,Ar-H),6.85(d,1H,J=8.6Hz,Ar-H),6.80(d,1H,J=8.6Hz,Ar-H),6.67(d,1H,J=8.6Hz,Ar-H),4.27(q,2H,J=7.1Hz,-CH2 -CH3),1.30(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.8,166.7,137.7,137.1,133.9,132.2,128.8,128.1,124.9,124.2,123.6,123.5,123.3,123.2,115.8,115.5×2,114.0,105.4,104.8,41.6,15.7.ESI-MS m/z 535.1/537.0/539.1[M+Na]+.
ii)2-(1-乙基-6-溴-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(96)的制备
按照化合物24c的制备方法,以化合物96a(100mg,0.195mmol)、HMDS(4ml,17.2mmol)和MeOH(0.5ml,8.6mmol)为原料制备,硅胶柱色谱分离、二氯甲烷洗脱得红色固体96(97mg,收率97%)。1H NMR(500MHz,DMSO-d6)δ11.82(s,1H,indole-NH),11.00(s,1H,imide-NH),7.79(d,1H,J=0.9Hz,Ar-H),7.78(s,1H,Ar-H),7.76(d,1H,J=7.5Hz,Ar-H),7.57(d,1H,J=7.6Hz,Ar-H),6.82(dd,1H,J=8.6Hz,1.6Hz,Ar-H),6.76(dd,1H,J=8.6Hz,1.7Hz,Ar-H),6.73(d,1H,J=8.5Hz,Ar-H),6.63(d,1H,J=8.6Hz,Ar-H),4.25(q,2H,J=7.2Hz,-CH2 -CH3),1.30(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ173.2×2,137.5,136.9,132.8,130.9,128.3,127.6,125.4,124.7,123.2,123.0,122.9,122.8,115.3,115.1,114.9,113.6,106.0,105.5,41.4,15.8.HR-ESIMS m/z 511.9617[M+H]+(calcd.for C22H16N3O2Br2,511.9609).
化合物97的制备
按照化合物2的制备方法,以化合物96(14mg,27.4μmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(12mg,137μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-6-溴-3-吲哚)-3-(1-羟甲基-6-溴-3-吲哚)马来酰亚胺(97)14.1mg,收率90%。1H NMR(500MHz,DMSO-d6)δ7.98(s,1H,Ar-H),7.82(s,1H,Ar-H),7.78(s,2H,Ar-H),6.85–6.82(m,2H,Ar-H),6.79(d,1H,J=8.8Hz,Ar-H),6.74(t,1H,J=7.5Hz,indole-CH2-OH),6.51(d,1H,J=8.5Hz,Ar-H),6.32(t,1H,J=6.0Hz,imide-CH2-OH),5.59(d,2H,J=7.5Hz,indole-CH2 -OH),4.95(d,2H,J=6.0Hz,imide-CH2 -OH),4.25(q,2H,J=6.8Hz,-CH2 -CH3),1.29(d,3H,J=6.8Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.3,171.2,137.0,136.9,133.4,133.0,127.8,127.2,125.5,125.4,123.4,123.3,123.2,123.0×2,115.4,114.5,113.8,105.6,105.4,69.8,60.9,41.4,15.8.HR-ESIMS m/z593.9650[M+Na]+(calcd.for C24H19N3O4Br2Na,593.9640).
化合物98的制备
按照化合物82的制备方法,以7-溴吲哚(850mg,5.6mmol)、Mg(134mg,5.6mmol)和化合物82a(286mg,1.1mmol)为原料,制得橙红色固体N-甲基-2,3-二(7-溴-3-吲哚)马来酰亚胺(98)180mg,收率32%。1H NMR(500MHz,DMSO-d6)δ11.94(s,2H,indole-NH),7.77(d,2H,J=2.7Hz,Ar-H),7.20(d,2H,J=7.5Hz,Ar-H),6.77(d,2H,J=8.0Hz,Ar-H),6.60(t,2H,J=7.8Hz,Ar-H),3.04(s,3H,-CH3).13C NMR(125MHz,DMSO-d6)δ171.8×2,134.7×2,130.5×2,127.7×2,127.4×2,124.9×2,121.3×2,120.6×2,107.0×2,104.8×2,24.5.HR-ESIMS m/z 497.9464[M+H]+(calcd.for C21H14N3O2Br2,497.9453).
化合物99的制备
按照化合物83的制备方法,以化合物98(317mg,0.638mmol)、NaH(28.1mg,0.702mmol,质量分数60%,分散于石蜡中)和EtI(57μL,0.702mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=5:1(v/v)洗脱得橙红色固体N-甲基-2-(1-乙基-7-溴-3-吲哚)-3-(7-溴-3-吲哚)马来酰亚胺(99)110mg,收率33%。1H NMR(500MHz,DMSO-d6)δ11.96(s,1H,indole-NH),7.83(s,1H,Ar-H),7.77(s,1H,Ar-H),7.24(d,1H,J=7.5Hz,Ar-H),7.20(d,1H,J=7.5Hz,Ar-H),6.95(d,1H,J=7.9Hz,Ar-H),6.64(t,1H,J=6.7Hz,Ar-H),6.62(d,1H,J=6.6Hz,Ar-H),6.57(t,1H,J=7.8Hz,Ar-H),4.59(q,2H,J=7.2Hz,-CH2 -CH3),3.04(s,3H,-NCH3),1.31(d,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.7×2,134.8×2,131.9,130.8,129.8,128.7,127.5,127.0,126.6,124.9,121.5,121.3,121.1,120.6,106.9,105.6,104.8,103.4,43.5,24.5,17.9.HR-ESIMS m/z525.9772[M+H]+(calcd.for C23H18N3O2Br2,525.9766).
化合物100的制备
i)2-(1-乙基-7-溴-3吲哚)-3-(7-溴-3-吲哚)马来酸酐(100a)的制备
按照化合物84a的制备方法,以化合物99(200mg,0.38mmol)为原料制备,硅胶柱色谱分离、二氯甲烷洗脱得橙红色固体100a(84mg,收率43%)。1H NMR(600MHz,DMSO-d6)δ12.23(s,1H,indole-NH),7.90(d,1H,J=2.5Hz,Ar-H),7.87(s,1H,Ar-H),7.29(d,1H,J=7.5Hz,Ar-H),7.26(d,1H,J=7.3Hz,Ar-H),7.00(d,1H,J=8.0Hz,Ar-H),6.71–6.68(m,1H,Ar-H),6.68–6.62(m,2H,Ar-H),4.60(q,2H,J=7.1Hz,-CH2 -CH3),1.32(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ166.6,166.5,135.9,135.1,132.2,130.0,129.5,128.1,127.8,126.7,125.5,122.1,122.0,121.4,121.0,106.4,105.2,104.9,103.7,43.8,17.9.ESI-MS m/z 512.9/514.9/516.9[M+H]+.
ii)2-(1-乙基-7-溴-3-吲哚)-3-(7-溴-3-吲哚)马来酰亚胺(100)的制备
按照化合物24c的制备方法,以化合物100a(150mg,0.293mmoll)、HMDS(4mL,17.2mmol)和MeOH(0.5mL,8.6mmol)为原料制备,硅胶柱色谱分离、二氯甲烷洗脱得红色固体100(123mg,
收率82%)。1H NMR(500MHz,DMSO-d6)δ11.94(s,1H,indole-NH),11.03(s,1H,imide-NH),7.80(s,1H,Ar-H),7.76(s,1H,Ar-H),7.23(d,1H,J=7.6Hz,Ar-H),7.19(d,1H,J=7.4Hz,Ar-H),6.92(d,1H,J=8.0Hz,Ar-H),6.64(d,1H,J=7.9Hz,Ar-H),6.62(t,1H,J=7.9Hz,Ar-H),6.56(t,1H,J=7.7Hz,Ar-H),4.58(d,2H,J=7.1Hz,-CH2 -CH3),1.32(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ172.9,172.8,134.8×2,131.9,130.8,129.9,129.2,127.5,127.2,127.1,124.8,121.4,121.3,121.1,120.6,106.9,105.6,104.8,103.3,43.5,17.9.HR-ESIMS m/z 511.9613[M+H]+(calcd.for C22H16N3O2Br2,511.9609).
化合物101的制备
按照化合物2的制备方法,以化合物100(55mg,107μmol)、甲醛溶液(3mL,质量分数37%)和NaHCO3(45mg,537μmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色固体N-羟甲基-2-(1-乙基-7-溴-3-吲哚)-3-(1-羟甲基-7-溴-3-吲哚)马来酰亚胺(101)20.2mg,收率33%。1H NMR(500MHz,DMSO-d6)δ12.06(s,1H,indole-NH),7.87(s,1H,),7.82(s,1H,Ar-H),7.26(d,2H,J=7.6Hz,Ar-H),7.22(d,1H,J=7.4Hz,Ar-H),6.95(d,1H,J=8.0Hz,Ar-H),6.65(d,1H,J=7.8Hz,Ar-H),6.63(d,1H,J=6.0Hz,Ar-H),5.82(t,1H,J=7.0Hz,-CH2OH),4.97(d,2H,J=7.0Hz,-CH2 OH),4.62(q,2H,J=7.0Hz,-CH2 -CH3),1.33(t,3H,J=7.0Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ170.5,170.4,134.3×2,134.2,131.3,130.4,129.2,128.2,126.9,126.9,126.4×2,126.1×2,120.9,120.8,119.9,104.8,104.2,60.1,42.9,17.2.HR-ESIMS m/z 541.9725[M+H]+(calcd.forC23H18N3O3Br2,541.9715).
化合物102的制备
i)N-甲基-2-(3-吲哚)-3-溴马来酰亚胺(102a)的制备
在50mL两口瓶中以THF(5mL)悬浮镁屑(360mg,15mmol),缓慢滴加溴乙烷(1.12mL,15mmol),室温搅拌20min后升温至45℃搅拌20min,滴加THF(5mL)溶解的吲哚(1.75g,15mmol),继续搅拌30min。降至室温,滴加THF(10mL)溶解的化合物82a(2g,7.5mmol),室温搅拌过夜。TLC检测至反应完毕,缓慢滴加饱和氯化铵水溶液(50mL)淬灭反应,乙酸乙酯萃取(2次×100mL),饱和食盐水洗(2次×100mL),合并有机相,并用无水硫酸钠干燥,真空旋蒸除去溶剂,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得橙色固体(102a)2.2g,收率96%。1H NMR(500MHz,DMSO-d6)δ12.12(s,1H,indole-NH),8.06(d,1H,J=8.0Hz,Ar-H),7.91(d,1H,J=8.0Hz,Ar-H),7.55(dt,1H,J=8.0Hz,0.6Hz,Ar-H),7.23(dt,1H,J=7.9Hz,0.9Hz,Ar-H),7.15(dt,1H,J=7.7Hz,1.0Hz,Ar-H),3.01(s,3H,-NCH3).13C NMR(125MHz,DMSO-d6)δ169.7,167.1,138.2,137.1,131.6,125.1,123.1,122.8,121.0,114.1,112.9,104.4,25.1.ESI-MS m/z 304.9[M+H]+.
ii)N-甲基-2-(1-叔丁基氧羰基-3-吲哚)-3-溴马来酰亚胺(102b)的制备
0℃下,在250mL单口瓶中,以THF(80mL)溶解化合物102a(2g,6.58mmol),加入催化量的DMAP,缓慢滴加THF(20mL)溶解的(Boc)2O(2.9g,13.16mmol),升至室温,搅拌反应2h。TLC检测至反应完毕,真空旋蒸除去溶剂,以硅胶柱色谱分离、石油醚:乙酸乙酯=10:1(v/v)洗脱得黄固体(102b)2.5g,收率94%。1H NMR(500MHz,DMSO-d6)δ8.13(d,1H,J=8.4Hz,Ar-H),8.08(s,1H,Ar-H),7.77(d,1H,J=7.9Hz,Ar-H),7.43(dt,1H,J=7.9Hz,0.9Hz,Ar-H),7.36(dt,1H,J=7.7Hz,1.0Hz,Ar-H),3.02(s,3H,-NCH3),1.65(s,9H,-C(CH3)3).13C NMR(125MHz,DMSO-d6)δ168.9,166.4,149.0,136.3,135.1,129.4,127.2,125.8,123.7,123.0,122.1,115.5,109.0,85.7,28.1,25.3.ESI-MS m/z 405.0[M+H]+.
iii)N-甲基-2-(1-叔丁基氧羰基-3-吲哚)-3-(3-吲哚)马来酰亚胺(102c)的制备
按照化合物102a的合成方法,由化合物102b(1.8g,4.46mmol)、镁屑(321mg,13.37mmol)、溴乙烷(1mL,13.37mmol)和吲哚(1.57g,13.37mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得黄色固体(102c)1.6g,收率81%。1H NMR(500MHz,DMSO-d6)δ11.83(s,1H,indole-NH),8.03(d,1H,J=8.3Hz,Ar-H),7.91(s,1H,Ar-H),7.85(d,1H,J=2.8Hz,Ar-H),7.37(d,1H,J=8.1Hz,Ar-H),7.18(t,1H,J=7.7Hz),6.98(t,1H,J=7.6Hz,Ar-H),6.87(d,1H,J=8.2Hz,Ar-H),6.86(d,1H,J=8.3Hz,Ar-H),6.82(t,1H,J=7.5Hz,1H,Ar-H),6.67(t,1H,J=7.6Hz,Ar-H),3.04(s,3H,-NCH3),1.60(s,9H,-C(CH3)3).13C NMR(125MHz,DMSO-d6)δ171.6,171.5,149.1,136.6,134.7,132.5,131.0,128.4,128.1,125.5,125.0,123.4,122.9,122.4,121.7,121.2,120.3,115.1,112.4,111.2,105.7,85.0,28.0,24.5.ESI-MS m/z 442.2[M+H]+.
iv)对甲基苯磺酸苯乙酯(102d)的制备
0℃下,在250mL两口瓶中以二氯甲烷(50mL)溶解苯乙醇(2g,16.3mmol),加入三乙胺(3.38mL,24.5mmol),滴加二氯甲烷(20mL)溶解的对甲苯磺酰氯(4.67g,24.5mmol),滴毕,升至室温,反应过夜。TLC检测至反应完毕,真空旋蒸除去溶剂,以硅胶柱色谱分离、石油醚:乙酸乙酯=25:1(v/v)洗脱得黄色固体(102d)3.6g,收率80%。ESI-MS m/z 277.1[M+H]+.
v)N-甲基-2-(1-叔丁基氧羰基-3-吲哚)-3-(1-苯乙基-3-吲哚)马来酰亚胺(102e)的制备
在25mL三口反应瓶中,以DMF(5mL)悬浮氢化钠(11mg,0.272mmol,含量60%分散在石蜡油中),–5℃搅拌30min后,缓慢滴加DMF(5mL)溶解的化合物102c(60mg,0.136mmol),低温继续搅拌45min,缓慢滴加DMF(2mL)溶解的化合物102d(100μL,0.272mmol),升至室温,反应过夜。TLC检测至反应完毕,降温至0℃,缓慢滴加饱和氯化铵水溶液(50mL)淬灭反应,乙酸乙酯萃取(3次×100mL),卤水洗(2次×100mL),合并有机相,并用无水硫酸钠干燥,真空旋蒸除去溶剂,硅胶柱色谱分离、石油醚:乙酸乙酯=7:1(v/v)洗脱得红色固体47mg,收率63%。1H NMR(500MHz,DMSO-d6)δ8.03(d,1H,J=8.4Hz,Ar-H),7.93(s,1H,Ar-H),7.72(s,1H,Ar-H),7.47(d,1H,J=8.3Hz,Ar-H),7.22(t,1H,J=6.6Hz,Ar-H),7.18(m,2H,Ar-H),7.14(m,2H,Ar-H),7.03–6.98(dt,1H,J=7.8Hz,0.9Hz,Ar-H),6.81(m,3H,Ar-H),6.66(dd,1H,J=7.0Hz,7.1Hz,Ar-H),4.47(t,2H,J=7.1Hz,N-CH2 -CH2Ph),3.01(s,3H,-NCH3),2.99(t,2H,J=7.1Hz,NCH2-CH2 -Ph),1.61(s,9H,-C(CH3)3).13C NMR(125MHz,DMSO-d6)δ171.5,171.4,149.2,138.7,136.2,134.7,133.7,131.7,129.2×2,128.7×2,128.6,128.2,126.9,126.1,125.1,123.3,122.8,122.6,121.7,121.4,120.5,115.1,111.2,110.9,104.9,85.1,47.8,36.1,28.1×3,24.5.ESI-MS m/z 546.3[M+H]+.
vi)N-甲基-2-(3-吲哚)-3-(1-苯乙基-3-吲哚)马来酰亚胺(102f)的制备
在100mL单口瓶中以甲苯(10mL)溶解化合物102e(34mg,0.062mmol),加入硅胶(200mg),加热回流2h。冷却至室温,TLC检测至反应完毕,真空旋蒸除去溶剂,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色固体(102f)26mg,收率94%。1H NMR(500MHz,DMSO-d6)δ11.68(s,1H,indole-NH),7.76(s,1H,Ar-H),7.63(s,1H,Ar-H),7.45(d,1H,J=8.3Hz,Ar-H),7.36(d,1H,J=8.1Hz,Ar-H),7.25(d,1H,J=7.8Hz,Ar-H),7.24(d,2H,J=7.2Hz,Ar-H),7.18–7.16(m,3H,Ar-H),6.99(d,1H,J=7.9Hz,Ar-H),6.95(d,1H,J=7.8Hz,Ar-H),6.75(d,1H,J=8.3Hz,Ar-H),6.73(d,1H,J=8.0Hz,Ar-H),6.64(d,1H,J=8.1Hz,Ar-H),6.60(d,1H,J=8.1Hz,Ar-H),4.46(t,2H,J=7.3Hz,N-CH2 -CH2Ph),3.03(t,2H,J=7.3Hz,NCH2-CH2 -Ph),3.01(s,3H,-NCH3).13C NMR(125MHz,DMSO-d6)δ172.2,172.1,138.8,136.4,136.0,134.7,132.3,129.6,129.2×2,128.7×2,127.4,126.9,126.7,126.4,125.8,122.1,121.6,121.4,119.9,119.7,112.2,110.6,106.1,105.3,47.7,36.2,24.4.ESI-MS m/z446.3[M+H]+.
vii)6-甲基-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(102)的制备
按照化合物73a的制备方法,由化合物102f(406mg,0.912mmol)、DDQ(269mg,1.19mmol)和p-TsOH(154mg,0.81mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色固体(102)340mg,收率84%。1H NMR(500MHz,DMSO-d6)δ11.83(s,1H,indole-NH),9.03(d,1H,J=8.0Hz,Ar-H),9.00(d,1H,J=7.8Hz,Ar-H),7.77(d,1H,J=8.1Hz,Ar-H),7.55(t,1H,J=7.5Hz,Ar-H),7.50(d,1H,J=8.1Hz,Ar-H),7.44(t,1H,J=7.0Hz,Ar-H),7.33(t,1H,J=6.9Hz,Ar-H),7.27(t,1H,J=6.9Hz,Ar-H),7.03–7.00(m,5H,Ar-H),5.07(t,2H,J=7.4Hz,N-CH2 -CH2Ph),3.05(s,3H,NCH3),3.03(t,2H,J=7.4Hz,NCH2-CH2 -Ph).13C NMR(125MHz,DMSO-d6)δ170.1×2,141.7,141.6,138.4,129.8,129.6,129.5,128.7×2,128.6,128.5,127.4,127.2,126.8,124.8,124.7,121.5,120.8,120.7×2,119.7,117.5,116.6,112.6,110.6,45.8,36.9,24.1.ESI-MS m/z 444.2[M+H]+.
化合物103的制备
i)12-苯乙基-12,13-二氢呋喃[3,4-c]吲哚[2,3-a]咔唑-5,7-二酮(103a)的制备
按照化合物73b的合成方法,由化合物102(200mg,0.45mmol)和KOH(5M,30mL)合成,得到黄色固体(103a)164mg,收率84%。由于产物的溶解性极差,且反应较完全,故未经分离纯化直接投入下一步反应。
ii)12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(103)的制备
按照化合物23c的合成方法,由化合物103a(20mg,0.047mmol),HMDS(500μL,2.35mmol)和甲醇(50μL,1.18mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色固体(103)16mg,收率90%。1H NMR(500MHz,DMSO-d6)δ11.83(s,1H,indole-NH),11.02(s,1H,imide-NH),9.08(d,1H,J=7.8Hz,Ar-H),9.06(d,1H,J=7.8Hz,Ar-H),7.79(d,1H,J=8.1Hz,Ar-H),7.62(d,1H,J=8.3Hz,Ar-H),7.56(t,1H,J=7.6Hz,Ar-H),7.47(t,1H,J=7.7Hz,Ar-H),7.35(t,1H,J=7.5Hz,Ar-H),7.30(t,1H,J=7.5Hz,Ar-H),7.10–6.98(m,5H,Ar-H),5.20(t,2H,J=7.1Hz,N-CH2 -CH2-Ph),3.07(t,2H,J=7.1Hz,N-CH2-CH2 -Ph).13C NMR(125MHz,DMSO-d6)δ171.6,171.5,141.6,141.5,138.2,133.0,129.5×2,128.9,128.4×2,127.3,127.1,126.7,124.9,124.7,123.5,121.6,121.4,120.7×2,120.3,117.4,116.6,112.5,110.6,45.8,36.5.ESI-MS m/z 430.2[M+H]+.
化合物104的制备
按照化合物2的制备方法,以化合物103(18mg,0.06mmol)为原料制备,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色荧光粉末6-羟甲基-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(104)21mg,收率80%。1H NMR(500MHz,DMSO-d6)δ11.92(s,1H,indole-NH),9.07(d,1H,J=7.9Hz,Ar-H),9.06(d,1H,J=7.8Hz,Ar-H),7.79(d,1H,J=8.1Hz,Ar-H),7.58(d,1H,J=8.2Hz,Ar-H),7.56(d,1H,J=8.1Hz,Ar-H),7.47(t,1H,J=7.6Hz,Ar-H),7.35(t,1H,J=7.4Hz,Ar-H),7.30(t,1H,J=7.4Hz,Ar-H),7.00–6.97(m,5H),6.29(t,1H,J=7.0Hz,-CH2OH),5.16(t,2H,J=6.8Hz,N-CH2 -CH2Ph),5.04(d,2H,J=7.0Hz,-CH2 OH),3.08(t,2H,J=6.8Hz,NCH2-CH2 -Ph).13C NMR(125MHz,DMSO-d6)δ169.5,169.4,141.6,141.5,138.2,130.0,129.4×2,128.9,128.4,127.4,127.2,126.7,124.9,124.7,124.5,123.5,121.4,121.3,120.8,120.7,119.1,117.4,116.6,112.6,110.6,60.2,45.8,36.4.ESI-MS m/z 460.2[M+H]+.
化合物105的制备
按照化合物14的合成方法,由化合物103a(40mg,0.09mmol)和乙二胺(75μL,0.9mmol)合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-氨乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(105)34mg,收率80%。1H NMR(500MHz,DMSO-d6)δ12.07(s,1H,indole-NH),9.08(d,1H,J=7.2Hz,Ar-H),9.07(d,1H,J=7.9Hz,Ar-H),7.83(d,1H,J=8.0Hz,Ar-H),7.62(d,1H,J=8.1Hz,Ar-H),7.58(t,1H,J=7.8Hz,Ar-H),7.47(t,1H,J=7.3Hz,Ar-H),7.35(t,1H,J=7.3Hz,Ar-H),7.30(t,1H,J=7.2Hz,Ar-H),7.01(m,5H,Ar-H),5.23(t,2H,J=6.6Hz,N-CH2 -CH2-Ph),3.90(t,2H,J=5.7Hz,N-CH2 -CH2NH2),3.10(t,2H,J=5.7Hz,NCH2-CH2 -NH2),3.09(t,2H,J=6.6Hz,N-CH2-CH2 -Ph).13C NMR(125MHz,DMSO-d6)δ170.2×2,141.7,141.6,138.2,129.5×2,128.8,128.4×2,127.4,127.2,126.7,124.7,124.6,121.9,121.4,120.9,120.7,119.4,119.3,117.5,116.7,112.7,111.6,110.7,45.9,40.9,40.9,36.5.ESI-MS m/z 473.3[M+H]+.
化合物106的制备
按照化合物16的合成方法,由化合物105(25mg,0.05mmol)制得黄色固体6-(2-氨乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮盐酸盐(106)173mg,收率90%。1H NMR(500MHz,DMSO-d6)δ12.24(s,1H,indole-NH),9.05(d,1H,J=7.5Hz,Ar-H),9.04(d,1H,J=7.0Hz,Ar-H),8.21(brs,3H,-NH3 +),7.87(d,1H,J=8.1Hz,Ar-H),7.56(d,1H,J=7.7Hz,Ar-H),7.55(d,1H,J=8.1Hz,Ar-H),7.45(d,1H,J=7.3Hz,Ar-H),7.34(t,1H,J=7.5Hz,Ar-H),7.28(t,1H,J=7.4Hz,Ar-H),7.03–6.93(m,5H),5.23(t,2H,J=6.4Hz,N-CH2 -CH2Ph),3.97(t,2H,J=6.4Hz,N-CH2 -CH2NH3 +),3.18(t,2H,J=6.4Hz,NCH2-CH2 -NH3 +),3.07(t,2H,J=6.4Hz,NCH2-CH2 -Ph).13C NMR(125MHz,DMSO-d6)δ170.1×2,141.8,141.6,138.2,129.8,129.4,128.7,128.3,127.4,127.1,126.6,124.6,124.5,121.4,120.9,121.2,120.7,120.6,119.3×2,117.4,116.7,112.8,110.7,110.6.45.8,38.2,36.6,35.7.ESI-MS m/z 473.2[M–Cl]+.
化合物107的制备
按照化合物14的合成方法,由化合物103a(25mg,0.06mmol)和1,3-丙二胺(99μL,1.2mmol)合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(3-氨丙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(107)22mg,收率75%。1H NMR(500MHz,DMSO-d6)δ12.19(s,1H,indole-NH),9.05(d,1H,J=7.8Hz,Ar-H),9.03(d,1H,J=7.8Hz,Ar-H),7.87(d,1H,J=8.1Hz,Ar-H),7.57(dt,1H,J=8.1Hz,1.1Hz,Ar-H),7.54(d,1H,J=8.1Hz,Ar-H),7.45(dt,1H,J=8.1Hz,1.1Hz,Ar-H),7.35(dt,1H,J=8.1Hz,1.1Hz,Ar-H),7.28(t,1H,J=7.9Hz,Ar-H),7.05–7.03(m,5H,Ar-H),5.20(t,2H,J=7.0Hz,N-CH2 -CH2Ph),3.76(t,2H,J=6.6Hz,N-CH2 -(CH2)2NH2),3.07(t,2H,J=7.0Hz,NCH2-CH2 -Ph),2.90(t,2H,J=6.6Hz,N(CH2)2-CH2 -NH2),2.04–2.00(m,2H,NCH2-CH2 -CH2NH2).13C NMR(125MHz,DMSO-d6)δ170.2×2,141.9,141.7,138.4,130.0,129.6×2,128.9,128.5×2,127.5,127.3,126.8,124.8,124.6,121.5,121.4,120.8×2,119.1×2,117.6,116.8,112.9,110.7,45.9,37.5,36.7,35.2,27.4.ESI-MS m/z 487.1[M+H]+.
化合物108的制备
按照化合物16的合成方法,由化合物107(23mg,0.05mmol)制得黄色固体6-(3-氨丙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮盐酸盐(108)20mg,收率90%。1H NMR(500MHz,DMSO-d6)δ12.18(s,1H,indole-NH),9.06(d,2H,J=7.9Hz,Ar-H),8.01(brs,3H,-NH3 +),7.85(d,1H,J=8.1Hz,Ar-H),7.59(d,1H,J=8.1Hz,Ar-H),7.56(d,1H,J=8.1Hz,Ar-H),7.45(t,1H,J=7.6Hz,Ar-H),7.34(t,1H,J=7.5Hz,Ar-H),7.29(t,1H,J=7.5Hz,Ar-H),7.04–6.97(m,5H),5.22(t,2H,J=6.8Hz,N-CH2 -CH2Ph),3.77(d,2H,J=6.8Hz,N-CH2 -(CH2)2NH3 +),3.07(t,2H,J=6.7Hz,NCH2-CH2 -Ph),2.93(t,2H,J=6.7Hz,N(CH2)2-CH2 -NH3 +),2.05–1.97(m,2H,NCH2-CH2 -CH2NH3 +).13C NMR(125MHz,DMSO-d6)δ170.2,170.1,141.8,141.6,138.3,130.0,129.8,129.5,128.8,128.4,127.4,127.3,127.2,126.7,124.7,124.5,121.4,121.2,120.7×2,119.0×2,117.5,116.7,112.8,110.7,45.8,37.4,36.5,35.1,27.3.ESI-MS m/z 487.1[M–Cl]+.
化合物109的制备
按照化合物24的合成方法,由化合物103a(21mg,0.049mmol)、4-(2-氨乙基)-吗啉(50μL,0.49mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-(4-吗啉)乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(109)19mg,收率80%。1H NMR(500MHz,DMSO-d6)δ11.91(s,1H,indole-NH),9.05(d,1H,J=7.9Hz,Ar-H),9.04(d,1H,J=7.8Hz,Ar-H),7.78(d,1H,J=8.1Hz),7.57(d,1H,J=7.8Hz,Ar-H),7.57(t,1H,J=7.3Hz,Ar-H),7.46(d,1H,J=7.2Hz,Ar-H),7.34(t,1H,J=7.2Hz,Ar-H),7.29(t,1H,J=7.4Hz,Ar-H),7.03–7.01(m,5H,Ar-H),5.16(t,2H,J=7.0Hz,N-CH2 -CH2Ph),3.78(t,2H,J=6.5Hz,imide-N-CH2 -CH2-morpholine),3.50(t,4H,J=3.9Hz,morpholine-N(CH2-CH 2)2O),3.07(t,2H,J=7.0Hz,NCH2-CH2 -Ph),2.61(t,2H,J=6.5Hz,imide-NCH2-CH2 -morpholine),2.47(t,4H,J=3.9Hz,morpholine-N(CH 2-CH2)2O).13C NMR(125MHz,DMSO-d6)δ170.0,169.9,141.6,141.5,138.2,130.0,129.8,129.5×2,128.8,128.7,128.4×2,127.4,127.2,126.7,124.6,121.4,121.3,120.8,120.1,119.0,117.4,116.7,112.6,110.6,66.6×2,56.5×2,53.6,45.8,36.5,34.9.ESI-MS m/z 543.2[M+H]+.
化合物110的制备
按照化合物24的合成方法,由化合物103a(23mg,0.054mmol)、4-(2-氨乙基)-哌嗪(60μL,0.54mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-哌嗪乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(110)16mg,收率80%。1H NMR(500MHz,DMSO-d6)δ12.12(s,1H,indole-NH),10.24(s,1H,piperazin-NH),9.04(d,1H,J=7.1Hz,Ar-H),9.03(d,1H,J=6.9Hz,Ar-H),7.83(d,1H,J=8.1Hz,Ar-H),7.58–7.52(m,2H,Ar-H),7.44(t,1H,J=7.5Hz,Ar-H),7.33(t,1H,J=7.4Hz,Ar-H),7.28(t,1H,J=7.4Hz,Ar-H),7.05–6.94(m,5H,Ar-H),5.19(t,2H,J=7.0Hz,N-CH2 -CH2Ph),3.80(t,2H,J=6.5Hz,imide-N-CH2 -CH2-piperazine),3.07(t,2H,J=7.0Hz,NCH2-CH2 -Ph),3.01(t,4H,J=4.4Hz,piperazine-N(CH 2-CH2)2NH),2.75(t,4H,J=4.4Hz,piperazine-N(CH2-CH 2)2NH),2.48(t,2H,J=6.5Hz,imide-NCH2-CH2 -piperazine).13C NMR(125MHz,DMSO-d6)δ170.0,169.9,141.7,141.6,138.2,129.9,129.5×2,128.7,128.4×2,127.4,127.1,126.7,124.7,124.5,121.4,121.2,120.7,120.6,119.0,118.9,117.4,116.6,112.7,110.6,55.6×2,53.6,49.5×2,43.1,36.5,34.8.ESI-MS m/z 542.3[M+H]+.
化合物111的制备
按照化合物24的合成方法,由化合物103a(15mg,0.035mmol)、2-氯-6-氟苯乙胺(20μL,0.35mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-氯-6-氟苯乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(111)13mg,收率68%。1H NMR(500MHz,DMSO-d6)δ11.93(s,1H,indole-NH),9.00(d,1H,J=7.8Hz,Ar-H),8.98(d,1H,J=7.9Hz,Ar-H),7.78(d,1H,J=8.1Hz,Ar-H),7.60(d,1H,J=8.4Hz,Ar-H),7.55(t,1H,J=7.6Hz,Ar-H),7.46(t,1H,J=7.4Hz,Ar-H),7.35(t,1H,J=7.7Hz,Ar-H),7.29(d,1H,J=7.6Hz,Ar-H),7.25(m,2H,Ar-H),7.11(dd,J=7.1Hz,5.9Hz,1H),7.08–6.95(m,5H,Ar-H),5.17(t,2H,J=7.0Hz,N-CH2 -CH2Ph),3.94(t,2H,J=6.5Hz,imide-N-CH2 -CH2-C6H3FCl),3.19(t,2H,J=6.5Hz,imide-NCH2-CH2 -C6H3FCl),3.07(t,2H,J=7.0Hz,NCH2-CH2 -Ph).13C NMR(125MHz,DMSO-d6)δ169.8,169.7,161.8(d,1JCF=244Hz),141.6(d,3JCF=11Hz),138.2,135.0,129.8(d,2JCF=11Hz),129.4×2,128.7,128.4×2,127.4,127.2,126.7,125.8×2,124.8,124.7,124.6,124.5×2,121.4(d,2JCF=22Hz),120.8,120.7,119.0,118.9,117.4,116.6,114.7(d,2JCF=23Hz),112.6,110.6,45.8,36.9,36.5,25.9.ESI-MS m/z 586.2[M+H]+.
化合物112的制备
按照化合物24的合成方法,由化合物103a(15mg,0.035mmol)、1-(2-氨基乙基)哌啶(50μL,0.47mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-哌啶乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(112)17mg,收率67%。1H NMR(500MHz,DMSO-d6)δ11.92(s,1H,indole-NH),9.04(d,1H,J=8.2Hz,Ar-H),9.02(d,1H,J=8.5Hz,Ar-H),7.78(d,1H,J=8.1Hz,Ar-H),7.56(d,1H,J=7.3Hz,Ar-H),7.54(d,1H,J=8.0Hz,Ar-H),7.45(d,1H,J=7.2Hz,Ar-H),7.34(d,1H,J=7.3Hz,Ar-H),7.28(d,1H,J=7.3Hz,Ar-H),7.08–6.96(m,5H,Ar-H),5.14(t,2H,J=6.6Hz,N-CH2 -CH2Ph),3.77(t,2H,J=5.4Hz,imide-N-CH2 -CH2-piperidine),3.06(t,2H,J=6.6Hz,NCH2-CH2 -Ph),2.65(t,2H,J=6.5Hz,imide-NCH2-CH2 -piperidine),2.49(t,4H,J=3.1Hz,piperidine-N(CH 2-CH2)2CH2),1.45(m,4H,piperidine-N(CH2-CH 2)2CH2),1.34(t,2H,J=3.4Hz,piperidine-N(CH2-CH2)2CH 2).13C NMR(125MHz,DMSO-d6)δ169.9×2,141.6,141.5,138.2,129.8,129.4×2,128.7,128.4×2,127.4,127.1,126.7,124.7,124.6,121.4,121.3,120.7,120.6,119.1,119.0,117.4,116.5,112.6,110.6,56.5×2,54.2,45.8,36.5,25.9,25.6×2,24.1.ESI-MS m/z 541.2[M+H]+.
化合物113的制备
按照化合物24的合成方法,由化合物103a(23mg,0.054mmol),4-甲基-1-哌嗪乙胺(50μL,0.55mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-(4-甲基哌嗪)乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(113)19mg,收率65%。1H NMR(500MHz,DMSO-d6)δ11.94(s,1H,indole-NH),9.06(d,1H,J=7.3Hz,Ar-H),9.05(d,1H,J=7.0Hz,Ar-H),7.79(d,1H,J=8.2Hz,Ar-H),7.59(d,1H,J=7.7Hz,Ar-H),7.56(d,1H,J=7.7Hz,Ar-H),7.46(t,1H,J=7.6Hz,Ar-H),7.35(t,1H,J=7.5Hz,Ar-H),7.07–6.97(m,5H,Ar-H),5.17(t,2H,J=6.8Hz,N-CH2 -CH2Ph),3.79(t,2H,J=6.3Hz,N-CH2 -CH2-piperazine),3.07(t,2H,J=6.9Hz,NCH2-CH2 -Ph),2.62(t,2H,J=6.5Hz,NCH2-CH2 -piperazine),2.48(t,4H,J=4.5Hz,piperazine-N(CH 2-CH2)2NCH3),2.35(t,4H,J=4.5Hz,piperazine-N(CH2-CH 2)2NCH3),2.15(s,3H,piperazine-N(CH2-CH2)2NCH 3).13C NMR(125MHz,DMSO-d6)δ169.9×2,141.6×2,138.2,129.9,129.5×2,128.7,128.4×2,127.5,127.2,126.7,124.7,124.6,121.4,121.3,120.8,120.7,119.1,119.0,117.4,116.7,112.6,110.6,55.9×2,54.8×2,52.6,45.8,45.6,36.5,35.2.ESI-MS m/z 556.3[M+H]+.
化合物114的制备
按照化合物24的合成方法,由化合物103a(40mg,0.093mmol),乙醇胺(102μL,1.86mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-羟乙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(114)28mg,收率67%。1H NMR(500MHz,DMSO-d6)δ11.86(s,1H,indole-NH),9.06(d,1H,J=8.0Hz,Ar-H),9.04(d,1H,J=7.9Hz,Ar-H),7.78(d,1H,J=8.1Hz,Ar-H),7.57(t,1H,J=7.3Hz,Ar-H),7.53(d,1H,J=8.1Hz,Ar-H),7.45(t,1H,J=7.3Hz,Ar-H),7.35(d,1H,J=7.0Hz,Ar-H),7.28(t,1H,J=7.5Hz,Ar-H),7.07–7.00(m,5H,Ar-H),5.12(t,2H,J=6.6Hz,N-CH2 -CH2Ph),4.93(t,1H,J=5.7Hz,-OH),3.72(t,2H,J=5.7Hz,N-CH2 -CH2OH),3.69(t,2H,J=5.7Hz,-CH2 OH),3.07(t,2H,J=6.6Hz,NCH2-CH2 -Ph).13C NMR(125MHz,DMSO-d6)δ170.2×2,141.7,141.6,138.4,129.8,129.5×2,128.8,128.5×2,127.5,127.2,126.8,124.9,124.8,121.5,121.4,120.8,120.7,119.3×2,117.5,116.7,112.6,110.6,58.9,45.9,45.8,36.6.ESI-MS m/z 474.3[M+H]+.化合物115的制备
按照化合物24的合成方法,由化合物103a(30mg,0.07mmol),3-羟基丙胺(100μL,1.4mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(3-羟丙基)-12-苯乙基-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(115)24mg,收率70%。1H NMR(500MHz,DMSO-d6)δ11.93(s,1H,indole-NH),9.07(d,1H,J=7.9Hz,Ar-H),9.05(d,1H,J=7.9Hz,Ar-H),7.80(d,1H,J=8.1Hz,Ar-H),7.58(dd,1H,J=8.3Hz,1.1Hz,Ar-H),7.57(t,1H,J=7.7Hz,Ar-H),7.48(dt,1H,J=8.1Hz,1.1Hz,Ar-H),7.35(d,J=8.3Hz,1H,Ar-H),7.29(t,1H,J=7.5Hz,Ar-H),7.08–7.01(m,5H,Ar-H),5.15(t,2H,J=7.1Hz,N-CH2 -CH2Ph),4.59(t,1H,J=5.1Hz,N(CH2)2CH2-OH),3.73(t,2H,J=7.2Hz,N-CH2 -CH2CH2OH),3.53–3.51(m,2H,N(CH2)2-CH2 -OH),3.07(t,2H,J=7.1Hz,NCH2-CH2 -Ph),1.88–1.83(m,2H,NCH2-CH2 -CH2OH).13C NMR(125MHz,DMSO-d6)δ170.2×2,141.8,141.7,138.4,129.9,129.6×2,128.8,128.5×2,127.5,127.3,126.9,124.9,124.8,121.6,121.4,120.8,120.7,119.2×2,117.5,116.8,112.9,110.7,59.3×2,45.9,36.6,32.3.ESI-MS m/z 488.1[M+H]+.
化合物116的制备
i)对甲苯磺酸(1-萘乙)酯(116a)的制备
按照化合物102d的合成方法,由萘乙醇(5g,0.029mmol),对甲苯磺酰氯(11.65g,0.061mmol)和三乙胺(8.46mL,0.061mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得白色固体9.6g,收率85%。ESI-MS m/z 278.1[M+H]+.
ii)N-甲基-2-(1-叔丁基氧羰基-3-吲哚)-3-(1-(1-萘乙基)-3-吲哚)马来酰亚胺(116b)的制备
按照化合物102e的合成方法,由化合物102c(1g,2.27mmol),氢化钠(200mg,4.54mmol)和化合物116a(1.13g,3.41mmol)合成。硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得红色固体(116b)563mg,收率41%。1H NMR(500MHz,DMSO-d6)δ8.08(d,1H,J=8.0Hz,Ar-H),8.07(d,1H,J=8.0Hz,Ar-H),7.97(s,1H,Ar-H),7.94(d,1H,J=7.8Hz,Ar-H),7.80(d,1H,J=8.2Hz,Ar-H),7.74(s,1H,Ar-H),7.59(dd,1H,J=8.3Hz,1.4Hz,Ar-H),7.56(d,1H,J=6.9Hz,Ar-H),7.44(d,1H,J=8.3H,Ar-H),7.38(t,1H,J=7.2Hz,Ar-H),7.24(d,1H,J=7.2Hz,Ar-H),7.19(d,1H,J=7.1Hz,Ar-H),7.04(t,1H,J=7.6Hz,Ar-H),6.95(d,1H,J=8.0Hz,Ar-H),6.89(d,1H,J=7.9Hz,Ar-H),6.85(d,1H,J=7.9Hz,Ar-H),6.74(t,1H,J=7.6Hz,Ar-H),4.58(t,2H,J=7.3Hz,N-CH2 -CH2-Nap),3.46(t,2H,J=7.3Hz,NCH2-CH2 -Nap),3.04(s,3H,NCH3),1.64(s,9H,-C(CH3)3).13C NMR(125MHz,DMSO-d6)δ171.5×2,149.3,136.4,134.9,134.8,134.0,133.9,131.9,131.8,129.3,128.4,128.3,127.8,127.4,126.9,126.3,125.9,125.2,124.0,123.6,123.0,122.8,122.0,121.7,120.7,115.3,111.3,110.9,105.1,85.2,47.2,33.3,28.2×3,24.9.ESI-MS m/z596.2[M+H]+.
iii)N-甲基-2-(3-吲哚)-3-(1-(1-萘乙基)-3-吲哚)马来酰亚胺(116c)的制备
按照化合物102f的合成方法,由化合物116b(100mg,0.168mmol)和硅胶(400mg)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得黄色固体(116c)79mg,收率95%。1HNMR(500MHz,DMSO-d6)δ11.72(d,1H,J=2.5Hz,indole-NH),8.09(d,1H,J=8.2Hz,Ar-H),7.94(s,1H,Ar-H),7.80(d,1H,J=8.2Hz,Ar-H),7.75(d,1H,J=2.8Hz,Ar-H),7.62(s,1H,Ar-H),7.58(t,1H,J=7.7Hz,Ar-H),7.54(t,1H,J=7.7Hz,Ar-H),7.43–7.40(m,2H,Ar-H),7.36(dd,1H,J=8.2Hz,7.0Hz,Ar-H),7.25(d,1H,J=7.0Hz,Ar-H),7.01(dt,2H,J=7.5Hz,1.1Hz,Ar-H),6.84(d,1H,J=8.0Hz,Ar-H),6.80(d,1H,J=8.1Hz,Ar-H),6.69(dt,1H,J=7.5Hz,1.1Hz,Ar-H),6.66(dt,1H,J=7.5Hz,1.1Hz,Ar-H),4.57(t,2H,J=7.3Hz,N-CH2 -CH2-Nap),3.51(t,2H,J=7.3Hz,NCH2-CH2 -Nap),3.02(s,3H,NCH3).13C NMR(125MHz,DMSO-d6)δ172.3,172.2,136.6,136.2,134.9,134.0,132.5,131.9,129.9,129.3,127.7,127.5,126.9,126.4,126.3,126.0,125.7,124.1,122.3×2,121.9,121.6,120.1,119.9,112.4,110.6,106.2,105.6,47.2,33.3,24.5.ESI-MS m/z 496.2[M+H]+.
iv)6-甲基-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(116)的制备
按照化合物102的合成方法,由化合物116c(400mg,0.808mmol)、DDQ(238mg,1.05mmol)和p-TsOH(154mg,0.81mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得黄色固体(116)270mg,收率65%。1H NMR(600MHz,DMSO-d6)δ11.85(s,1H,indole-NH),9.07(d,1H,J=7.9Hz,Ar-H),9.00(d,1H,J=7.8Hz,Ar-H),8.00(d,1H,J=7.9Hz,Ar-H),7.83(d,1H,J=7.3Hz,Ar-H),7.73(d,1H,J=8.1Hz,Ar-H),7.62(d,1H,J=8.1Hz,Ar-H),7.60(dt,1H,J=7.7Hz,1.1Hz,Ar-H),7.48–7.40(m,2H,Ar-H),7.39(t,1H,J=7.7Hz,Ar-H),7.33(t,1H,J=7.7Hz,Ar-H),7.23(d,1H,J=7.7Hz,Ar-H),7.22(d,1H,J=8.1Hz,Ar-H),7.16(t,1H,J=7.4Hz,Ar-H),7.02(d,1H,J=6.6Hz,Ar-H),5.24(t,2H,J=7.0Hz,N-CH2 -CH2-Nap),3.53(t,2H,J=7.0Hz,NCH2-CH2 -Nap),3.12(s,3H,NCH3).13C NMR(150MHz,DMSO-d6)δ170.2,170.1,141.8,141.7,134.5,133.8,132.0,129.9,129.0,128.8,127.6,127.5×2,127.0,126.6,126.1,125.7,124.8,124.7,123.8,121.6×2,120.9,120.7,119.4,119.3,117.7,117.0,112.6,110.2,45.3,33.4,24.1.ESI-MS m/z 494.2[M+H]+.
化合物117的制备
i)12-(1-萘乙基)-12,13-二氢呋喃[3,4-c]吲哚[2,3-a]咔唑-5,7-二酮(117a)的制备
按照化合物73b的合成方法,由化合物116(200mg,0.45mmol)和KOH(5M,30mL)合成,得到黄色固体(117a)164mg,收率84%。由于产物的溶解性极差,且反应较完全,故未经分离纯化直接投入下一步反应。
ii)12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(117)的制备
按照化合物23c的合成方法,由化合物117a(26mg,0.054mmol)、HMDS(500μL,2.35mmol)和甲醇(50μL,1.18mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得黄色固体(117)22mg,收率87%。1H NMR(500MHz,DMSO-d6)δ11.91(s,1H,indole-NH),11.03(s,1H,imide-NH),9.10(d,1H,J=7.9Hz,Ar-H),9.04(d,1H,J=7.8Hz,Ar-H),8.00(d,1H,J=8.0Hz,Ar-H),7.80(d,1H,J=7.3Hz,Ar-H),7.75(d,1H,J=7.5Hz,Ar-H),7.62(d,1H,J=8.1Hz,Ar-H),7.60(t,1H,J=7.8Hz,Ar-H),7.47–7.42(m,2H,Ar-H),7.37(t,1H,J=7.7Hz,Ar-H),7.33–7.30(m,2H,Ar-H),7.25(d,1H,J=7.6Hz,Ar-H),7.15(t,1H,J=7.8Hz,Ar-H),7.05(d,1H,J=7.0Hz,Ar-H),5.32(t,2H,J=7.0Hz,N-CH2 -CH2-Nap),3.54(t,2H,J=7.0Hz,NCH2-CH2 -Nap).13C NMR(125MHz,DMSO-d6)δ171.7,171.6,141.8,134.5,133.8,132.0,130.2,129.1,129.0,127.7,127.6,127.5,127.1,126.6,126.4,126.1,125.7,125.0,124.9,123.8,121.8,121.7,120.9,120.8,120.5,120.4,117.6,117.0,112.6,110.3,45.4,33.3.ESI-MS m/z 480.3[M+H]+.
化合物118的制备
按照化合物2的合成方法,由化合物117(20mg,0.04mmol)和甲醛(3mL,质量分数37%)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得黄色固体6-羟甲基-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(118)16mg,收率80%。1H NMR(600MHz,DMSO-d6)δ11.90(s,1H,imide-NH),9.08(d,1H,J=7.9Hz,Ar-H),9.00(d,1H,J=7.4Hz,Ar-H),7.96(d,1H,J=7.6Hz,Ar-H),7.77(d,1H,J=8.4Hz,Ar-H),7.72(d,1H,J=8.2Hz,Ar-H),7.59(d,1H,J=8.1Hz,Ar-H),7.56(d,J=7.4Hz,1H),7.45–7.39(m,2H),7.36(t,1H,J=7.5Hz,Ar-H),7.29(t,1H,J=7.8Hz,Ar-H),7.22(t,2H,J=7.0Hz,Ar-H),7.10(t,1H,J=7.6Hz,Ar-H),6.98(d,1H,J=6.9Hz,Ar-H),6.30(t,1H,J=6.6Hz,-CH2OH),5.26(t,2H,J=6.9Hz,N-CH2 -CH2-Nap),5.03(d,2H,J=6.6Hz,-CH2 OH),3.53(t,2H,J=6.9Hz,NCH2-CH2 -Nap).13C NMR(150MHz,DMSO-d6)δ169.4,169.3,141.7,141.6,134.3,133.6,131.9,130.1,128.9×2,127.5,127.4,127.0,126.4,126.3,126.0,125.5,124.6,124.5,123.7,122.3,121.5,121.4,120.8,120.6,119.1,119.0,117.5,116.8,112.6,110.2,60.3,45.3,33.2.ESI-MS m/z 510.2[M+H]+.
化合物119的制备
按照化合物14的合成方法,由化合物117a(36mg,0.074mmol)和乙二胺(500μL,7.5mmol)合成,硅胶柱色谱分离、二氯甲烷:甲醇=3:1(v/v)洗脱得黄色固体6-(2-氨乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(119)35mg,收率90%。1H NMR(500MHz,DMSO-d6)δ9.10(d,1H,J=7.9Hz,Ar-H),9.02(d,1H,J=7.7Hz,Ar-H),8.02(dd,1H,J=6.7Hz,2.2Hz,Ar-H),7.82(dd,1H,J=6.4Hz,2.5Hz,Ar-H),7.81(d,1H,J=8.3Hz,Ar-H),7.62(d,1H,J=8.2Hz,Ar-H),7.61(t,1H,J=7.6Hz,Ar-H),7.48(dd,2H,J=6.6Hz,3.2Hz,Ar-H),7.40(t,1H,J=7.7Hz,Ar-H),7.31(t,1H,J=8.1Hz,Ar-H),7.23(t,1H,J=8.2Hz,Ar-H),7.20(d,1H,J=7.8Hz,Ar-H),7.11(t,1H,J=8.1Hz,Ar-H),6.96(d,1H,J=6.9Hz,Ar-H),5.33(t,2H,J=6.1Hz,N-CH2 -CH2-Nap),3.93(t,2H,J=6.7Hz,N-CH2 -CH2NH2),3.54(t,2H,J=6.1Hz,NCH2-CH2 -Nap),3.14(t,2H,J=6.7Hz,NCH2-CH2 -NH2).13C NMR(125MHz,DMSO-d6)δ170.3,170.2,142.0,141.9,134.5,133.8,132.0,130.0,129.1,129.0,127.7,127.6,127.1,126.6,126.4,126.1,125.7,124.8,124.7,123.8,121.6,121.5,121.0,120.7,119.6,119.5,117.7,117.1,112.9,110.3,45.4,39.1,37.1,33.5.ESI-MS m/z 523.4[M+H]+.
化合物120的制备
按照化合物16的合成方法,由化合物119(23mg,0.04mmol)和盐酸的乙酸乙酯溶液(3N,3mL)合成得20mg黄色固体6-(2-氨乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮盐酸盐(120),收率90%。1H NMR(500MHz,DMSO-d6)δ12.16(s,1H,indole-NH),9.07(d,1H,J=7.9Hz,Ar-H),9.00(d,1H,J=7.6Hz,Ar-H),8.13(s,3H,-NH3 +),8.00(d,1H,J=5.6Hz,Ar-H),7.79(d,2H,J=7.7Hz,Ar-H),7.59(d,1H,J=8.0Hz,Ar-H),7.56(d,1H,J=7.5Hz,Ar-H),7.45(d,1H,J=7.5Hz,Ar-H),7.43(d,1H,J=7.5Hz,Ar-H),7.36(t,1H,J=7.4Hz,Ar-H),7.28(t,1H,J=7.6Hz,Ar-H),7.22(d,1H,J=7.4Hz,Ar-H),7.18(t,1H,J=6.8Hz,Ar-H),7.07(t,1H,J=7.5Hz,Ar-H),6.92(d,1H,J=6.9Hz,Ar-H),5.33(t,2H,J=6.9Hz,N-CH2 -CH2-Nap),3.97(t,2H,J=6.7Hz,N-CH2 -CH2NH3 +),3.52(t,2H,J=6.9Hz,NCH2-CH2 -Nap),3.19(t,2H,J=6.7Hz,NCH2-CH2 -NH3 +).13C NMR(125MHz,DMSO-d6)δ170.1,170.0,141.8×2,134.3,133.6,131.8,129.8,129.0,128.8,127.5,127.4,127.0,126.5,126.0,125.5,124.6,124.5,123.7,121.4,121.3,120.8,120.6,119.4,119.3,117.6,116.9,112.8,110.2,45.3,38.3,35.7,33.4.ESI-MS m/z523.3[M–Cl]+.
化合物121的制备
按照化合物14的合成方法,由化合物117a(25mg,0.05mmol)、1,3-丙二胺(60μL,0.78mmol)合成得25mg黄色固体6-(3-氨丙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(121),收率90%。1H NMR(500MHz,DMSO-d6)δ12.16(s,1H,indole-NH),9.08(d,1H,J=8.0Hz,Ar-H),9.01(d,1H,J=7.7Hz,Ar-H),7.97(d,1H,J=7.4Hz,Ar-H),7.81–7.79(m,2H,Ar-H),7.62(t,1H,J=7.5Hz,Ar-H),7.61(d,1H,J=7.6Hz,Ar-H),7.59(t,1H,J=7.5Hz,Ar-H),7.48–7.42(m,2H,Ar-H),7.37(t,1H,J=7.4Hz,Ar-H),7.31(t,1H,J=7.6Hz,Ar-H),7.23(t,1H,J=7.7Hz,Ar-H),7.21(t,1H,J=7.4Hz,Ar-H),7.11(t,1H,J=7.8Hz,Ar-H),6.99(d,1H,J=7.0Hz,Ar-H),5.33(t,2H,J=6.8Hz,N-CH2 -CH2-Nap),3.79(t,2H,J=7.2Hz,N-CH2 -(CH2)2-NH2),3.54(t,2H,J=6.8Hz,NCH2-CH2 -Nap),2.91(t,2H,J=7.2Hz,N(CH2)2-CH 2-NH2),2.11–1.93(m,2H,NCH2-CH2 -CH2NH2).13C NMR(125MHz,DMSO-d6)δ170.3,170.2,142.0,141.9,134.5,133.8,132.0,130.1,129.0×2,127.7,127.6,127.5,127.1,126.6,126.1,125.7,124.8,124.7,123.8,121.6,121.5,120.9,120.7,119.2,119.1,117.7,117.1,112.9,110.4,45.9,45.4,37.6,35.2,27.4.ESI-MS m/z 537.4[M+H]+.
化合物122的制备
按照化合物16的合成方法,由化合物121(25mg,0.04mmol)和盐酸的乙酸乙酯溶液(3N,3mL)合成得黄色固体6-(3-氨丙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮盐酸盐(122)20mg,收率90%。1H NMR(500MHz,DMSO-d6)δ12.14(s,1H,indole-NH),9.07(d,1H,J=7.8Hz,Ar-H),9.01(d,1H,J=7.8Hz,Ar-H),7.99(s,3H,-NH3 +),7.96(d,1H,J=9.0Hz,Ar-H),7.78(d,2H,J=7.9Hz,Ar-H),7.59(d,1H,J=8.0Hz,Ar-H),7.56(d,1H,J=7.9Hz,Ar-H),7.46–7.40(m,2H,Ar-H),7.35(t,1H,J=7.5Hz,Ar-H),7.28(d,1H,J=7.2Hz,Ar-H),7.24(d,1H,J=7.9Hz,Ar-H),7.22(d,1H,J=7.2Hz,Ar-H),7.11(t,1H,J=7.5Hz,Ar-H),6.98(d,1H,J=6.8Hz,Ar-H),5.34(t,2H,J=6.8Hz,N-CH2 -CH2-Nap),3.81(t,2H,J=7.2Hz,N-CH2 -(CH2)2-NH3 +),3.52(t,2H,J=6.8Hz,NCH2-CH2 -Nap),2.93(t,2H,J=7.2Hz,N(CH2)2-CH2 -NH3 +),2.05–1.99(m,2H,NCH2-CH2 -CH2NH2).13C NMR(125MHz,DMSO-d6)δ170.1,170.0,141.8,141.7,134.3,133.6,131.8,130.0,129.3,128.9×2,127.5,127.4,127.0,126.4,125.9,125.5,124.6,124.5,123.6,121.4,121.3,120.8,120.6,119.1,119.0,117.6,116.9,112.7,110.3,45.3,37.4,35.1,33.3,27.3.ESI-MS m/z573.3[M–Cl]+.
化合物123的制备
按照化合物24的合成方法,由化合物117a(20mg,0.04mmol),4-(2-氨乙基)-吗啉(50μL,0.42mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=3:1(v/v)洗脱得黄色固体6-(2-吗啉乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(123)19mg,收率76%。1H NMR(500MHz,DMSO-d6)δ11.90(s,1H,indole-NH),9.07(d,1H,J=8.0Hz,Ar-H),9.00(d,1H,J=7.8Hz,Ar-H),7.97(d,1H,J=7.7Hz,Ar-H),7.79(dd,1H,J=6.9Hz,2.4Hz,Ar-H),7.72(d,1H,J=8.1Hz,Ar-H),7.60(d,1H,J=8.2Hz,Ar-H),7.57(t,1H,J=7.7Hz,Ar-H),7.47–7.40(m,2H,Ar-H),7.36(t,1H,J=7.5Hz,Ar-H),7.29(d,1H,J=6.9Hz,Ar-H),7.25(d,1H,J=8.6Hz,Ar-H),7.23(t,1H,J=7.3Hz,Ar-H),7.14(t,1H,J=7.7Hz,Ar-H),7.02(d,1H,J=6.9Hz,Ar-H),5.28(t,2H,J=6.4Hz,N-CH2 -CH2-Nap),3.80(t,2H,J=5.8Hz,N-CH2 -CH2-morpholine),3.54(t,2H,J=6.4Hz,NCH2-CH2 -Nap),3.56(t,4H,J=3.9Hz,morpholine-N(CH2-CH 2)2O),2.63(t,2H,J=5.8Hz,imide-NCH2-CH2 -morpholine),2.49(t,4H,J=3.9Hz,morpholine-N(CH 2-CH2)2O).13C NMR(125MHz,DMSO-d6)δ169.9,169.8,141.7,134.4,133.6,131.9,129.9,128.9×2,127.5,127.4,127.0,126.4,126.0,125.6,124.7,124.6,123.7,121.5,121.4,120.8,120.6,119.1×2,117.6,116.9,112.5,110.2,66.6×2,56.5,53.6×2,45.3,34.9,33.2.ESI-MS m/z593.3[M+H]+.
化合物124的制备
按照化合物24的合成方法,由化合物117a(20mg,0.042mmol)、4-(2-氨乙基)哌嗪(45μL,0.42mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得黄色固体6-(2-哌嗪乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(124)20mg,收率80%。1H NMR(500MHz,DMSO-d6)δ9.05(d,1H,J=7.9Hz,Ar-H),8.97(d,1H,J=7.6Hz,Ar-H),7.94(d,1H,J=8.0Hz,Ar-H),7.84(d,1H,J=8.0Hz,Ar-H),7.80(d,1H,J=7.8Hz,Ar-H),7.59(d,1H,J=8.7Hz,Ar-H),7.57(d,1H,J=7.6Hz,Ar-H),7.46–7.39(m,2H,Ar-H),7.35(dd,1H,J=11.4Hz,4.3Hz,Ar-H),7.23(dd,1H,J=8.1Hz,1.2Hz,Ar-H),7.20(t,1H,J=7.6Hz,Ar-H),7.11(d,1H,J=8.2Hz,Ar-H),7.10(t,1H,J=8.2Hz,Ar-H),6.98(d,1H,J=6.9Hz,Ar-H),5.33(t,2H,J=6.6Hz,N-CH2 -CH2-Nap),3.78(t,2H,J=6.5Hz,N-CH2 -CH2-piperazine),3.56(t,2H,J=6.6Hz,NCH2-CH2 -Nap),2.97(t,4H,J=5.1Hz,piperazine-N(CH 2-CH2)2NH),2.70(t,4H,J=5.1Hz,piperazine-N(CH2-CH 2)2NH),2.68(t,2H,J=6.5Hz,NCH2-CH2 -piperazine).13C NMR(125MHz,DMSO-d6)δ170.1,170.0,142.0,141.8,134.6,134.3,133.7,133.3,131.9,131.5,130.0,129.0×2,128.1,127.6,127.0,126.5,126.0,125.6,124.7,123.8,121.5,120.8,120.6,119.2,119.0,117.6,116.9,112.9,110.2,55.9×2,50.0,45.4×2,43.5,35.0,33.5.ESI-MS m/z 592.3[M+H]+.
化合物125的制备
按照化合物24的合成方法,由化合物117a(20mg,0.042mmol)、2-氯-6-氟苯乙胺(30μL,0.42mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=25:1(v/v)洗脱得黄色固体6-(2-氯-6-氟苯乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(125)12mg,收率60%。1H NMR(500MHz,DMSO-d6)δ11.90(s,1H,indole-NH),9.02(d,1H,J=7.8Hz,Ar-H),8.95(d,2H,J=7.8Hz,Ar-H),7.95(d,1H,J=8.2Hz,Ar-H),7.80(d,1H,J=7.9Hz,Ar-H),7.73(d,1H,J=8.1Hz,Ar-H),7.62(d,1H,J=8.1Hz,Ar-H),7.58(t,1H,J=7.5Hz,Ar-H),7.47(t,1H,J=7.2Hz,Ar-H),7.42(dd,1H,J=8.2Hz,1.8Hz,Ar-H),7.36(t,1H,J=7.5Hz,Ar-H),7.31(d,1H,J=7.3Hz,Ar-H),7.29–7.27(m,2H,Ar-H),7.23(t,1H,J=7.0Hz,Ar-H),7.17(d,1H,J=7.1Hz,Ar-H),7.15(t,1H,J=7.0Hz,Ar-H),7.14(dd,1H,J=7.4Hz,1.2Hz,Ar-H),7.07(d,1H,J=6.9Hz,Ar-H),5.29(t,2H,J=6.8Hz,N-CH2 -CH2-Nap),3.95(t,2H,J=6.7Hz,N-CH2 -CH2-C6H3FCl),3.54(t,2H,J=6.8Hz,NCH2-CH2 -Nap),3.21(t,2H,J=6.7Hz,NCH2-CH2 -C6H3FCl).13C NMR(125MHz,DMSO-d6)δ169.9,169.8,162.7(d,1JCF=244Hz),141.8(d,3JCF=11Hz),135.1,134.8,134.5,133.7,133.4,132.0,130.1,129.9(d,3JCF=11Hz),128.9,128.8,127.6,127.5,127.1,126.5,126.1,125.9,125.7,124.9,124.8,123.8,122.8,121.6(d,2JCF=22Hz),120.9,120.8,119.1,119.0,117.7,117.0,114.9(d,2JCF=23Hz),112.6,110.3,90.3,45.4,36.5,33.3,29.4.ESI-MS m/z636.2[M+H]+.
化合物126的制备
按照化合物24的合成方法,由化合物117a(20mg,0.042mmol)、1-(2-氨基乙基)哌啶(50μL,0.42mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得黄色固体6-(2-哌啶乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(126)19mg,收率67%。1H NMR(500MHz,DMSO-d6)δ11.89(s,1H,indole-NH),9.05(d,1H,J=7.9Hz,Ar-H),8.97(d,1H,J=7.6Hz,Ar-H),7.95(d,1H,J=8.9Hz,Ar-H),7.78(d,1H,J=7.4Hz,Ar-H),7.71(d,1H,J=8.1Hz,Ar-H),7.59(d,1H,J=8.2Hz,Ar-H),7.56(t,1H,J=7.6Hz,Ar-H),7.44–7.40(m,2H,Ar-H),7.34(t,1H,J=7.5Hz,Ar-H),7.26(d,1H,J=7.0Hz,Ar-H),7.20(d,1H,J=8.0Hz,Ar-H),7.18(d,1H,J=7.5Hz,Ar-H),7.13(t,1H,J=7.5Hz,Ar-H),7.00(d,1H,J=6.8Hz,Ar-H),5.29(t,2H,J=6.8Hz,N-CH2 -CH2-Nap),3.73(t,2H,J=6.6Hz,N-CH2 -CH2-piperidine),3.51(t,2H,J=6.8Hz,NCH2-CH2 -Nap),2.58(t,2H,J=6.2Hz,NCH2-CH2 -piperidine),2.44(t,4H,J=3.3Hz,piperidine-N(CH 2-CH2)2CH2),1.45–1.41(m,4H,piperidine-N(CH2-CH 2)2CH2),1.33(t,2H,J=4.4Hz,piperidine-N(CH2-CH2)2CH 2).13C NMR(125MHz,DMSO-d6)δ169.9,169.8,141.7,141.6,135.1,134.4,133.6,131.9,129.9,128.9,128.8,127.5,127.4,127.4,126.9,126.4,126.0,125.6,125.1,124.7,124.6,123.7,121.4,121.4,120.7,120.6,119.1,119.0,117.5,116.9,112.5,110.1,56.6×2,54.3,45.2,35.2,33.3,25.9×2,24.3.ESI-MSm/z 591.3[M+H]+.
化合物127的制备
按照化合物24的合成方法,由化合物117a(22mg,0.046mmol)、4-甲基-1-哌嗪乙胺(50μL,0.55mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得黄色固体6-(2-(4-甲基哌嗪)乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(127)22mg,收率80%。1H NMR(500MHz,DMSO-d6)δ11.95(s,1H,indole-NH),9.06(d,1H,J=7.0Hz,Ar-H),8.98(d,1H,J=7.2Hz,Ar-H),7.95(d,1H,J=6.7Hz,Ar-H),7.78(d,1H,J=6.8Hz,Ar-H),7.73(d,1H,J=8.1Hz,Ar-H),7.62–7.54(m,2H,Ar-H),7.42(d,2H,J=7.8Hz,Ar-H),7.34(t,1H,J=7.8Hz,Ar-H),7.27(t,1H,J=7.3Hz,Ar-H),7.20(d,2H,J=6.9Hz,Ar-H),7.11(d,1H,J=7.6Hz,Ar-H),7.01(d,1H,J=7.6Hz,Ar-H),5.27(t,2H,J=6.3Hz,N-CH2 -CH2-Nap),3.75(t,2H,J=6.3Hz,N-CH2 -CH2-piperazine),3.51(t,2H,J=6.3Hz,NCH2-CH2 -Nap),2.60(d,2H,J=5.8Hz,NCH2-CH2 -piperazine),2.48(t,4H,J=4.6Hz,piperazine-N(CH 2-CH2)2NCH3),2.25(t,4H,J=4.6Hz,piperazine-N(CH2-CH 2)2NCH3),2.07(s,3H,piperazine-N(CH2-CH2)2NCH3 ).13C NMR(125MHz,DMSO-d6)δ169.9,169.8,141.7,141.6,134.4,133.6,131.8,129.9,128.9,127.5,127.4,126.9,126.4,125.9,125.6,124.7,124.6,123.7,121.4,121.4,120.8,120.6,119.15,119.0,117.5,116.9,112.6,110.1,56.1,55.1×2,53.0×2,46.1,45.3,35.2,33.3.ESI-MS m/z 605.4[M+H]+.
化合物128的制备
按照化合物24的合成方法,由化合物117a(30mg,0.063mmol)、乙醇胺(55μL,0.94mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(2-羟乙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(128)26mg,收率89%。1H NMR(500MHz,DMSO-d6)δ11.95(s,1H,indole-NH),9.11(d,1H,J=7.8Hz,Ar-H),9.05(d,1H,J=7.9Hz,Ar-H),8.02(d,1H,J=7.7Hz,Ar-H),7.83(d,1H,J=8.1Hz,Ar-H),7.75(d,1H,J=8.1Hz,Ar-H),7.63(d,1H,J=8.0Hz,Ar-H),7.59(t,1H,J=7.8Hz,Ar-H),7.47(m,2H,Ar-H),7.38(t,1H,J=7.5Hz,Ar-H),7.34(t,1H,J=7.8Hz,Ar-H),7.26–7.24(m,2H,Ar-H),7.15(t,1H,J=7.8Hz,Ar-H),7.00(d,1H,J=7.2Hz,Ar-H),5.32(t,2H,J=7.0Hz,N-CH2 -CH2-Nap),4.94(t,1H,J=6.1Hz,N-CH2 -CH2-OH),4.2(brs,-OH),3.80(t,2H,J=6.1Hz,NCH2-CH2 -OH),3.56(t,2H,J=7.0Hz,NCH2-CH2 -Nap).13CNMR(125MHz,DMSO-d6)δ170.3,170.2,141.9,134.5,133.8,132.0,130.1,129.1,129.0,127.7,127.6,127.5,127.1,126.6,126.1,125.7,124.9,124.8,123.8,121.6,121.6,120.9,120.8,119.8,119.4,117.7,117.0,112.7,112.4,110.3,58.9,45.4,40.7,33.4.ESI-MS m/z 524.4[M+H]+.
化合物129的制备
按照化合物24的合成方法,由化合物117a(25mg,0.052mmol)、3-羟基丙胺(50μL,0.52mmol)和催化量的三乙胺合成,硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得黄色固体6-(3-羟丙基)-12-(1-萘乙基)-12,13-二氢-5H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7(6H)-二酮(129)24mg,收率80%。1H NMR(500MHz,DMSO-d6)δ11.91(s,1H,indole-NH),9.10(d,1H,J=7.9Hz,Ar-H),9.04(d,1H,J=7.7Hz,Ar-H),8.01(d,1H,J=8.0Hz,Ar-H),7.82(dd,2H,J=7.6Hz,3.2Hz,Ar-H),7.74(d,1H,J=8.1Hz,Ar-H),7.62(d,1H,J=8.2Hz,Ar-H),7.59(dt,1H,J=8.2Hz,1.2Hz,Ar-H),7.48–7.43(m,2H,Ar-H),7.39(dt,1H,J=7.8Hz,1.1Hz,Ar-H),7.34(t,1H,J=7.8Hz,Ar-H),7.27(d,1H,J=8.1Hz,Ar-H),7.27(t,1H,J=7.6Hz,Ar-H),7.14(t,1H,J=7.7Hz,Ar-H),7.03(d,1H,J=6.6Hz,Ar-H),5.30(t,2H,J=6.9Hz,N-CH2 -CH2-Nap),4.57(brs,-OH),3.77(t,2H,J=7.2Hz,N-CH2 -CH2CH2OH),3.54(t,2H,J=6.9Hz,NCH2-CH2 -Nap),3.53(t,2H,J=6.2Hz,N(CH2)2-CH2 -OH),1.90–1.84(m,2H,NCH2-CH2 -CH2OH).13C NMR(125MHz,DMSO-d6)δ170.2,170.1,141.8,134.5,133.8,132.0,130.1,129.1,129.0,127.7,127.6,127.1,126.6,126.1,125.7,124.9,124.8,123.8,121.7,121.6,120.9,120.8,119.3,119.3,117.7,117.0,112.7,112.4,110.3,59.3,45.4,35.5,33.3,32.3.ESI-MS m/z 538.5[M+H]+.
化合物130的制备
氩气保护下,在25mL两口反应瓶中,加入十字孢碱(46.6mg,0.1mmol),用3mL二氯甲烷溶解,滴加过量的三乙胺,然后加入对氟苯磺酰氯,室温搅拌反应4h,加水终止反应,二氯甲烷萃取,并用无水Na2SO4干燥,真空蒸干,后经过凝胶柱色谱分离、甲醇洗脱得3′-N-对氟苯磺酰十字孢碱(130)52.0mg,收率83.3%。1H NMR(600MHz,CDCl3)δ9.40(d,1H,J=7.4Hz,Ar-H),7.92(m,2H,Ar-H),7.85(d,1H,J=7.3Hz,Ar-H),7.72(d,1H,J=7.8Hz,Ar-H),7.47(t,1H,J=7.3Hz,Ar-H),7.46(t,1H,J=7.3Hz,Ar-H),7.43(d,2H,J=7.3Hz,Ar-H),7.32(m,2H,J=7.3Hz,Ar-H),7.06(d,1H,J=7.8Hz,Ar-H),6.84(brs,1H,-NH),6.56(m,1H,H-1'),4.95(d,1H,J=16.0Hz,H-7a),4.89(d,1H,J=16.0Hz,H-7b),4.53(dd,1H,J=12.4Hz,5.5Hz,H-3'),3.94(s,1H,H-4'),2.71(s,3H,4'-OCH3),2.47(s,3H,3'-NCH3),2.43(m,1H,H-1'a),2.37(s,3H,H-6'),2.27(m,1H,H-2'b);13C NMR(150MHz,CDCl3)δ173.5,164.6(d,1JC-F=247.2Hz),138.4,136.5,136.5,132.5,130.5,129.7×2(d,3JC-F=9.2Hz),126.9,126.3,125.5,125.3,124.8,123.6,121.7,120.8,120.2,119.2,116.8×2(d,2JC-F=13.7Hz),116.3,114.6,112.2,107.6,94.7,86.5,82.4,60.3,52.1,46.0,30.8,29.1,28.3.ESI-MS m/z 625.3[M+H]+。
化合物131的制备
按照化合物130的合成方法,由十字孢碱(46.6mg,0.1mmol)、三乙胺和对氯苯磺酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得3′-N-对氯苯磺酰十字孢碱(131)51.3mg,收率80.1%。1H NMR(600MHz,CDCl3)δ9.41(d,1H,J=7.7Hz,Ar-H),7.86(d,1H,J=7.8Hz,Ar-H),7.84(d,2H,J=8.2Hz,Ar-H),7.72(d,1H,J=8.7Hz,Ar-H),7.61(d,2H,J=8.2Hz,Ar-H),7.48(t,1H,J=8.3Hz,Ar-H),7.44(t,1H,J=6.8Hz,Ar-H),7.36(t,1H,J=7.8Hz,Ar-H),7.34(t,1H,J=7.8Hz,Ar-H),7.09(d,1H,J=8.2,Ar-H),6.61(br s,1H,-NH),6.58(dd,1H,J=9.2Hz,4.1Hz,H-1'),4.93(m,2H,H-7),4.52(ddd,1H,J=12.8Hz,5.5Hz,1.9Hz,H-3'),3.96(s,1H,H-4'),2.72(s,3H,4'-OCH3),2.48(s,1H,3'-NCH3),2.44(m,1H,H-2'a),2.41(s,3H,H-6'),2.26(m,1H,H-2'a).13C NMR(150MHz,CDCl3)δ173.3,139.8,138.5,137.7,136.5,132.5,130.5,129.9×2,128.5×2,126.9,126.3,125.6,125.3,124.8,123.7,121.7,120.8,120.3,119.3,116.4,114.7,112.2,107.6,94.7,86.6,82.4,60.4,52.2,45.9,30.8,29.2,28.3.ESI-MS m/z 641.4/643.4[M+H]+。
化合物132的制备
按照化合物130的合成方法,由十字孢碱(46.6mg,0.1mmol)、三乙胺和对溴苯磺酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得3′-N-对溴苯磺酰十字孢碱(132)52.6mg,收率76.8%。1H NMR(600MHz,CDCl3)δ9.40(d,1H,J=7.1Hz,Ar-H),7.99(d,1H,J=6.9Hz,Ar-H),7.83(d,1H,J=7.7Hz,Ar-H),7.72(d,1H,J=8.3Hz,Ar-H),7.61(d,2H,J=6.9Hz,Ar-H),7.47(t,1H,J=7.7Hz,Ar-H),7.42(t,1H,J=7.2Hz,Ar-H),7.34(t,2H,J=7.1Hz,Ar-H),7.04(d,2H,J=7.7,Ar-H),6.51(br s,1H,-NH),4.99(d,1H,J=18.0Hz,H-7a),4.92(d,1H,J=16.4Hz,H-7b),4.50(dd,1H,J=5.5Hz,12.7Hz,H-3'),3.91(s,1H,H-4'),2.69(s,3H,4'-OCH3),2.47(s,1H,3'-NCH3),2.45(t,1H,J=12.6Hz,H-2'a),2.35(s,3H,H-6'),2.45(t,1H,J=12.4Hz,H-2'a);13C NMR(150MHz,CDCl3)δ173.4,138.4,132.9×2,138.3,136.4,132.5,130.5,128.6×2,128.2,126.8,126.3,125.5,125.3,124.7,123.6,121.7,120.7,120.3,119.0,116.2,114.6,112.2,107.7,94.6,86.6,82.4,60.3,52.2,46.1,30.8,29.1,28.3.ESI-MS m/z 685.3/687.3[M+H]+。
化合物133的制备
按照化合物130的合成方法,由十字孢碱(46.6mg,0.1mmol)、三乙胺和苯磺酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得3′-N-苯磺酰十字孢碱(133)50.4mg,收率83.2%。1HNMR(600MHz,CDCl3)δ9.40(d,1H,J=7.8Hz,Ar-H),7.91(d,2H,J=7.4Hz,Ar-H),7.87(d,1H,J=7.3Hz,Ar-H),7.70(d,1H,J=7.8Hz,Ar-H),7.64(t,2H,J=7.8Hz,Ar-H),7.46(t,1H,J=7.8Hz,Ar-H),7.42(t,1H,J=7.8Hz,Ar-H),7.34(t,1H,J=7.3Hz,Ar-H),7.32(t,1H,J=7.3Hz,Ar-H),7.07(d,1H,J=8.2Hz,Ar-H),6.71(br s,1H,-NH),6.69(dd,1H,J=9.2Hz,4.6Hz,H-1'),4.94(m,2H,H-7),4.54(dd,2H,J=12.8Hz,5.5Hz,H-3'),3.91(s,1H,H-4'),2.73(s,3H,4'-OCH3),2.45(s,3H,3'-NCH3),2.43(br m,1H,H-2'a),2.39(s,3H,H-6'),2.25(br m,1H,H-2'b).13C NMR(150MHz,CDCl3)δ173.3,139.1,138.4,136.4,133.3,132.4,130.4,129.5×2,127.0×2,126.8,126.2,125.4,125.1,124.7,123.6,121.5,120.6,120.2,119.1,116.2,114.5,112.1,107.5,94.6,86.2,82.4,60.3,51.9,45.9,30.7,29.1,28.1.ESI-MS m/z 607.3[M+H]+。
化合物134的制备
氩气保护下,在25mL两口反应瓶中,加入3'-N-对氟苯甲酰十字孢碱(15.0mg,0.025mmol),用1mL甲醇溶解,然后加入溴代丁二酰亚胺(5.0mg,0.027mmol),室温搅拌反应2h,加水终止反应,二氯甲烷萃取,并用无水Na2SO4干燥,真空蒸干,后经过凝胶柱色谱分离、甲醇洗脱得3-溴-3′-N-对氟苯甲酰十字孢碱(134)8.4mg,收率49.0%。1H NMR(600MHz,CDCl3)δ9.51(s,1H,Ar-H),7.86(d,1H,J=7.8Hz,Ar-H),7.75(d,1H,J=7.3Hz,Ar-H),7.46(t,2H,J=7.8Hz,Ar-H),7.43(m,2H,J=7.3Hz,Ar-H),7.36(t,1H,J=7.8Hz,Ar-H),7.34(d,1H,J=7.8Hz,Ar-H),7.09(d,2H,J=7.3Hz,Ar-H),7.02(br s,1H,-NH),6.66(br s,1H,H-1'),5.18(d,1H,J=7.8Hz,H-3'),4.93(br s,2H,H-7),4.17(s,1H,H-4'),2.82(s,3H,4'-OCH3),2.71(dt,1H,J=12.5Hz,3.2Hz,H-2'a),2.56(s,3H,3'-NCH3),2.44(dt,1H,J=12.5Hz,3.2Hz,H-2'b),2.36(s,3H,H-6').13C NMR(150MHz,CDCl3)δ171.6,171.4,163.5(d,1JC-F=247.2Hz),138.5×2,132.9,132.1,131.5,130.4,129.4×2(d,3JC-F=6.9Hz),129.0,128.1,126.8,125.4,125.1,124.6,121.8,120.8,119.2,115.7×2(d,2JC-F=13.7Hz),115.5,115.0,112.3,109.1,94.7,84.7,82.6,60.5,49.8,46.2,34.6,29.2,28.1.ESI-MS m/z 667.3/669.3[M+H]+。
化合物135的制备
i)化合物135a的制备
在充入氧气的25mL两口反应瓶中,加入十字孢碱(6.6mg,0.1mmol),用3mL DMSO溶解,加入过量的叔丁醇钾,室温反应6h,加水终止反应,乙酸乙酯萃取三次,真空蒸干,后经过凝胶柱色谱分离、甲醇洗脱得7-氧亚基十字孢碱(135a)42.5mg,收率88.6%。1H NMR(600MHz,DMSO-d6)δ11.04(s,1H,-NH),9.21(d,1H,J=8.8Hz,Ar-H),9.08(d,1H,J=7.7Hz,Ar-H),8.03(d,1H,J=8.8Hz,Ar-H),7.71(t,1H,J=8.8Hz,Ar-H),7.60(t,1H,J=7.7Hz,Ar-H),7.51(t,1H,J=7.7Hz,Ar-H),7.41(t,1H,J=7.7Hz,Ar-H),7.33(d,1H,J=7.7Hz,Ar-H),6.77(br s,1H,H-1'),4.14(m,1H,H-3'),3.38(s,1H,H-4'),3.36(s,3H,4'-OCH3),2.55(m,2H,H-2'),2.34(s,1H,3'-NCH3),1.22(s,3H,H-6').13C NMR(150MHz,DMSO-d6)δ171.4,171.2,142.1,139.2,132.3,131.7,127.5,126.5,126.4,125.6,124.4,122.8,121.2,121.1,120.6,120.2,117.2,116.4,115.8,109.2,92.1,84.5,81.3,57.4,51.5,33.6,30.4,30.1.ESI-MS m/z481.1[M+H]+。
ii)化合物135的制备
按照化合物130的合成方法,由135a(10.0mg,0.021mmol)、三乙胺和对氟苯甲酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得7-氧亚基-3′-N-对氟苯甲酰十字孢碱(135)10.8mg,收率86.1%。1H NMR(600MHz,CDCl3)δ9.33(d,1H,J=7.7Hz,Ar-H),9.17(d,1H,J=7.8Hz,Ar-H),7.73(d,1H,J=7.7Hz,Ar-H),7.68(t,1H,J=7.4Hz,Ar-H),7.56(t,1H,J=7.8Hz,Ar-H),7.51(t,1H,J=7.8Hz,Ar-H),7.42(t,1H,J=7.8Hz,Ar-H),7.41(d,2H,J=7.7Hz,Ar-H),7.41(t,1H,J=7.8Hz,Ar-H),7.11(d,2H,J=7.4Hz,Ar-H),6.72(br s,1H,H-1'),5.20(d,1H,J=5.8Hz,H-3'),4.16(s,1H,H-4'),2.86(s,3H,4'-OCH3),2.80(m,1H,H-2'a),2.53(s,3H,3'-NCH3),2.38(m,1H,H-2'b),2.35(s,3H,H-6').13C NMR(150MHz,CDCl3)δ169.8,169.6,163.8(d,1JC-F=270.0Hz),139.3,139.2,137.8,133.2×2,130.3,129.6×2(d,3JC-F=6.9Hz),127.0×2(d,2JC-F=13.7Hz),126.5,126.3,123.8,122.5,121.4,121.3,121.0,119.4,117.4,116.3,114.7,111.3,108.2,94.7,86.0,82.5,60.1,51.9,30.7,28.9,28.3.ESI-MS m/z 603.2[M+H]+。
化合物136的制备
按照化合物130的合成方法,由135a(10.0mg,0.021mmol)、三乙胺和对氯苯甲酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得7-氧亚基-3′-N-对氯苯甲酰十字孢碱(136)10.6mg,收率82.3%。1H NMR(600MHz,CDCl3)δ9.38(d,1H,J=7.7Hz,Ar-H),9.24(d,1H,J=7.8Hz,Ar-H),7.94(t,1H,J=7.4Hz,Ar-H),7.76(t,1H,J=7.8Hz,Ar-H),7.61(t,1H,J=7.8Hz,Ar-H),7.54(d,2H,J=7.7Hz,Ar-H),7.52(d,2H,J=7.7Hz,Ar-H),7.51(br s,1H,-NH),7.41(t,1H,J=7.8Hz,Ar-H),7.40(d,1H,J=7.8Hz,Ar-H),7.11(d,1H,J=7.4Hz,Ar-H),6.80(br s,1H,H-1'),5.26(dd,1H,J=5.4Hz,12.3Hz,H-3'),4.22(s,1H,H-4'),2.88(s,3H,4'-OCH3),2.83(dt,1H,J=3.2Hz,10.8Hz,H-2'a),2.74(dt,1H,J=3.2Hz,10.8Hz,H-2'b),2.58(s,3H,3'-NCH3),2.44(s,3H,H-6').13C NMR(150MHz,CDCl3)δ171.4,169.9,169.7,139.5,137.9,136.3,131.6,130.2,129.0,128.6,129.6×2,127.1×2,126.5,126.4,123.8,122.6,121.4,121.3,121.0,119.5,117.4,116.4,111.7,111.6,108.3,94.8,84.7,82.5,60.5,49.7,30.4,29.0,28.1.ESI-MS m/z619.5/621.5[M+H]+。
化合物137的制备
按照化合物130的合成方法,由135a(10.0mg,0.021mmol)、三乙胺和对溴苯甲酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得7-氧亚基-3′-N-对溴苯甲酰十字孢碱(137)11.1mg,收率80.3%。1H NMR(500MHz,CDCl3)δ9.34(d,1H,J=7.7Hz,Ar-H),9.19(d,1H,J=7.8Hz,Ar-H),7.74(t,1H,J=7.4Hz,Ar-H),7.71(t,1H,J=7.8Hz,Ar-H),7.56(t,1H,J=7.8Hz,Ar-H),7.54(d,2H,J=7.7Hz,Ar-H),7.52(d,2H,J=7.7Hz,Ar-H),7.51(br s,1H,-NH),7.41(t,1H,J=7.8Hz,Ar-H),7.40(d,1H,J=7.8Hz,Ar-H),7.29(d,1H,J=7.4Hz,Ar-H),6.71(br s,1H,H-1'),5.21(dd,1H,J=5.5Hz,12.4Hz,H-3'),4.15(s,1H,H-4'),2.84(s,3H,4'-OCH3),2.80(dt,1H,J=3.2Hz,10.8Hz,H-2'a),2.67(dt,1H,J=3.2Hz,10.8Hz,H-2'b),2.53(s,3H,3'-NCH3),2.35(s,3H,H-6').13C NMR(150MHz,CDCl3)δ171.4,170.0,169.7,139.4,137.8,134.8,131.5,130.1,129.0,128.6,128.7×2,127.1×2,126.5,126.3,126.2,123.8,122.5,121.4,121.3,121.1,119.5,117.4,116.3,111.6,108.3,94.7,84.7,82.5,60.4,49.7,34.4,28.9,28.1.ESI-MS m/z663.5/665.5[M+H]+。
化合物138的制备
按照化合物130的合成方法,由135a(10.0mg,0.021mmol)、三乙胺和对碘苯甲酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得7-氧亚基-3′-N-对碘苯甲酰十字孢碱(138)11.6mg,收率78.5%。1H NMR(600MHz,CDCl3)δ9.38(d,1H,J=7.7Hz,Ar-H),9.24(d,1H,J=7.8Hz,Ar-H),7.94(t,1H,J=7.4Hz,Ar-H),7.76(t,1H,J=7.8Hz,Ar-H),7.61(t,1H,J=7.8Hz,Ar-H),7.54(d,2H,J=7.7Hz,Ar-H),7.52(t,2H,J=7.7Hz,Ar-H),7.51(br s,1H,-NH),7.41(t,1H,J=7.8H,Ar-H),7.40(d,1H,J=7.8Hz,Ar-H),7.11(d,1H,J=7.4Hz,Ar-H),6.80(dd,1H,J=3.7Hz,9.2Hz,H-1'),5.26(dd,1H,J=5.5Hz,12.4Hz,H-3'),4.22(s,1H,H-4'),2.88(s,3H,4'-OCH3),2.83(dt,1H,J=3.2Hz,10.8Hz,H-2'a),2.74(dt,1H,J=3.2Hz,10.8Hz,H-2'b),2.58(s,3H,3'-NCH3),2.44(s,3H,H-6').13C NMR(150MHz,CDCl3)δ171.4,170.0,169.8,139.5,137.9,136.3,131.9,130.2,129.0,128.6,129.6×2,127.1×2,126.5,126.4,123.8,122.6,121.4,121.3,121.0,120.9,119.5,117.4,116.4,111.7,108.3,94.8,84.7,82.5,60.5,49.7,30.4,29.0,28.1.ESI-MS m/z 711.5[M+H]+.
化合物139的制备
氩气保护下,在25mL两口反应瓶中,加入3'-N-对氟苯甲酰十字孢碱(20.0mg,0.034mmol),用1mL DMSO溶解,然后加入0.03mL的2N NaOH溶液,室温搅拌反应4h,加水终止反应,乙酸乙酯萃取,并用无水Na2SO4干燥,真空蒸干,后经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-对氟苯甲酰十字孢碱(139)7.6mg,收率37.0%。1H NMR(600MHz,CDCl3)δ9.00(d,1H,J=7.7Hz,Ar-H),8.64(d,1H,J=7.7Hz,-NH),7.50(t,1H,J=8.2Hz,Ar-H),7.49(t,2H,J=8.2Hz,Ar-H),7.44(t,2H,J=7.3Hz,Ar-H),7.37(t,1H,J=7.7Hz,Ar-H),7.36(t,1H,J=7.7Hz,Ar-H),7.25(d,1H,J=7.7Hz,Ar-H),7.19(t,1H,J=7.7Hz,Ar-H),7.11(t,2H,J=7.7Hz,Ar-H),7.10(s,1H,H-7),6.71(br s,1H,-OH),6.47(br s,1H,H-1'),5.19(d,1H,J=8.2Hz,H-3'),3.84(s,1H,H-4'),2.70(s,3H,4'-OCH3),2.35(m,1H,H-2'a),2.24(m,1H,H-2'b),2.19(s,3H,3'-NCH3),2.04(s,3H,H-6').13C NMR(150MHz,CDCl3)δ173.3,171.5,163.6(d,1JC-F=247.2Hz),136.5×2,132.5,132.2,130.6,129.5×2(d,3JC-F=6.9Hz),126.7,126.4,125.5,125.2,124.9,123.8,121.6,120.6,120.3,119.2,116.3,115.8×2(d,2JC-F=13.7Hz),114.6,112.4,107.6,94.7,84.8,82.5,79.4,60.5,49.8,34.5,29.2,28.2.ESI-MS m/z 605.3[M+H]+.
化合物140和141的制备
按照化合物139的合成方法,由3'-N-对氯苯甲酰十字孢碱(20.0mg,0.034mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-对氯苯甲酰十字孢碱(140)7.2mg(收率35.1%)和7β-羟基-3′-N-对氯苯甲酰十字孢碱(141)5.4mg(收率26.3%)。
化合物140:1H NMR(600MHz,DMSO-d6)δ9.25(d,1H,J=7.8Hz,Ar-H),8.87(s,1H,-NH),8.46(d,1H,J=7.7Hz,Ar-H),7.99(d,1H,J=7.8Hz,Ar-H),7.68(d,2H,J=7.8Hz,Ar-H),7.52(d,2H,J=7.3Hz,Ar-H),7.50(t,1H,J=8.0Hz,Ar-H),7.49(t,2H,J=7.3Hz,Ar-H),7.34(t,1H,J=7.8Hz,Ar-H),7.31(t,1H,J=7.8Hz,Ar-H),7.19(s,1H,-OH),6.54(d,1H,J=10.0Hz,H-7),6.44(d,1H,J=8.0Hz,H-1'),5.19(d,1H,J=9.2Hz,H-3'),4.50(s,1H,H-4'),2.84(s,3H,4'-OCH3),2.73(m,1H,H-2'a),2.67(m,1H,H-2'b),2.54(s,3H,3'-NCH3),2.48(s,3H,H-6').13C NMR(150MHz,DMSO-d6)δ171.0,171.1,137.4,136.6,134.8,133.8,132.5,130.6,129.1×2,128.6×2,126.9,126.5,125.6,125.2,124.8,123.7,121.6,120.7,120.3,119.3,116.4,114.7,112.5,109.6,95.1,84.7,82.7,79.1,60.6,49.8,34.5,29.9,27.0.ESI-MS m/z605.3/607.3[M+H]+。
化合物141:1H NMR(600MHz,CDCl3)δ9.25(d,1H,J=7.8Hz,Ar-H),8.46(d,1H,J=7.7Hz,-NH),7.72(d,1H,J=7.8Hz,Ar-H),7.49(t,1H,J=7.8Hz,Ar-H),7.46(t,2H,J=7.8Hz,Ar-H),7.40(d,2H,J=7.3Hz,Ar-H),7.38(d,2H,J=7.3Hz,Ar-H),7.35(t,2H,J=7.8Hz,Ar-H),7.22(d,1H,J=7.8Hz,Ar-H),6.68(s,1H,H-7),6.66(s,1H,-OH),6.41(d,1H,J=8.0Hz,H-1'),5.19(d,1H,J=9.2Hz,H-3'),4.21(s,1H,H-4'),2.81(s,3H,4'-OCH3),2.71(m,1H,H-2'a),2.61(m,1H,H-2'b),2.51(s,3H,3'-NCH3),1.66(s,3H,H-6').13C NMR(150MHz,CDCl3)δ171.3,171.2,136.9,136.4,133.4,132.5,130.9,130.6,129.0×2,128.6×2,126.9,126.5,125.6,125.2,124.8,123.7,121.6,120.7,120.3,119.3,115.9,115.6,112.5,107.8,94.7,84.7,82.4,79.4,60.7,49.8,34.5,29.3,28.1.ESI-MS m/z 605.3/607.3[M+H]+。
化合物142和143的制备
按照化合物139的合成方法,由3'-N-对三氟甲基苯甲酰十字孢碱(20.0mg,0.031mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-对三氟甲基苯甲酰十字孢碱(142)7.5mg(收率36.6%)和7β-羟基-3′-N-对三氟甲基苯甲酰十字孢碱(143)5.4mg(收率26.3%)。
化合物142:1H NMR(600MHz,CDCl3)δ8.93(d,1H,J=7.4Hz,Ar-H),8.62(d,1H,J=7.4Hz,-NH),7.69(m,3H,Ar-H),7.60(d,1H,J=7.9Hz,Ar-H),7.51(m,2H,Ar-H),7.43(t,2H,J=7.8Hz,Ar-H),7.37(t,2H,J=7.8Hz,Ar-H),7.16(d,1H,J=7.8Hz,Ar-H),6.89(s,1H,H-7),6.87(m,1H,H-1'),6.49(br s,1H,-OH),5.19(m,1H,H-3'),3.90(s,1H,H-4'),2.70(s,3H,4'-OCH3),2.54(m,1H,H-2'a),2.35(m,1H,H-2'b),2.09(s,3H,3'-NCH3),2.06(s,3H,H-6').13C NMR(150MHz,CDCl3)δ170.2,170.1,141.1,139.7,138.3,134.8×2,132.8,130.1,128.8×2(q,2JC-F=27.0Hz),127.9,127.1,127.3,126.7,125.8,125.3(q,3JC-F=8.0Hz),125.1,124.6,122.7(q,1JC-F=270.0Hz),121.8,120.1(q,3JC-F=7.1Hz),119.6,115.0,114.7,113.2,109.0,94.5,83.0,82.0,78.2,60.3,48.8,33.7,29.3,28.2.ESI-MS m/z 655.2[M+H]+。
化合物143:1H NMR(600MHz,DMSO-d6)δ9.28(d,1H,J=7.4Hz,Ar-H),8.88(s,1H,-NH),8.51(d,1H,J=6.6Hz,Ar-H),7.99(d,1H,J=7.7Hz,Ar-H),7.83(d,1H,J=7.1Hz,Ar-H),7.70(d,1H,J=7.3Hz,Ar-H),7.66(d,1H,J=7.3Hz,Ar-H),7.49(m,3H,Ar-H),7.34(t,1H,J=8.3Hz,Ar-H),7.30(m,2H,Ar-H),7.09(d,1H,J=9.0Hz,H-7),6.46(br s,1H,-OH),6.45(m,1H,H-1'),5.09(d,1H,J=10.0Hz,H-3'),4.53(s,1H,H-4'),2.90(s,3H,4'-OCH3),2.78(m,1H,H-2'a),2.73(s,3H,3'-NCH3),2.57(m,1H,H-2'b),2.39(s,3H,H-6').13C NMR(150MHz,DMSO-d6)δ170.9,170.8,139.8,137.1,138.3,134.8×2,132.8,130.1,128.7×2(q,2JC-F=27.0Hz),127.9,127.1,127.3,126.7,125.8,125.0(q,3JC-F=8.0Hz),125.1,124.6,122.6(1JC-F=270.0Hz),121.8,120.1(q,3JC-F=7.1Hz),119.6,115.0,114.7,113.2,109.0,94.5,83.0,82.0,78.3,60.3,48.9,33.8,29.2,28.1.ESI-MS m/z 655.2[M+H]+。
化合物144和145的制备
按照化合物139的合成方法,由130(25.0mg,0.040mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-对氟苯磺酰十字孢碱(144)8.4mg(收率32.8%)和7β-羟基-3′-N-对氟苯磺酰十字孢碱(145)6.0mg(收率23.4%)。
化合物144:1H NMR(600MHz,DMSO-d6)δ9.22(d,1H,J=7.9Hz,Ar-H),8.86(s,1H,-NH),8.45(d,1H,J=8.6Hz,Ar-H),9.07(m,2H,Ar-H),7.99(d,1H,J=8.4Hz,Ar-H),7.56(m,2H,Ar-H),7.50(m,3H,Ar-H),7.34(t,1H,J=7.3Hz,Ar-H),7.30(d,1H,J=7.7Hz,H-7),6.95(m,1H,H-1'),6.44(br s,1H,-OH),4.53(d,1H,J=13.4Hz,H-3'),4.18(s,1H,H-4'),2.63(s,3H,4'-OCH3),2.59(s,3H,3'-NCH3),2.57(m,1H,H-2'a),2.43(m,1H,H-2'b),2.40(s,3H,H-6').13C NMR(150MHz,DMSO-d6)δ170.3,163.5(d,1JC-F=85.7Hz),138.8,136.5,134.9,130.2×2(d,3JC-F=9.9Hz),129.6,126.0,125.6,125.6,125.4,123.5,123.4,123.4,122.6,120.3,119.7,118.8,117.0×2(d,2JC-F=19.1Hz),115.1,114.8,113.0,109.1,94.8,85.1,82.1,60.3,51.8,40.1,30.60,29.0,26.9.ESI-MS m/z 641.2[M+H]+。
化合物145:1H NMR(600MHz,CDCl3)δ9.19(d,1H,J=8.2Hz,Ar-H),8.37(d,1H,J=7.9Hz,-NH),7.87(m,2H,Ar-H),7.65(d,1H,J=8.3Hz,Ar-H),7.44(ddd,1H,J=1.3,7.3,8.0Hz,Ar-H),7.36(ddd,1H,J=1.5,6.9,8.1Hz,Ar-H),7.31(d,1H,J=7.3Hz,Ar-H),7.29(dt,2H,J=2.8,8.2Hz,Ar-H),7.26(m,1H,Ar-H),7.24(t,1H,J=7.4Hz,Ar-H),6.99(d,1H,J=8.1Hz,Ar-H),6.82(s,1H,H-7),6.48(dd,1H,J=4.5,8.9Hz,H-1'),6.27(s,1H,-OH),4.49(ddd,1H,J=2.1,5.5,13.0Hz,H-3'),3.87(s,1H,H-4),2.57(s,3H,4'-OCH3),2.44(s,3H,3'-NCH3),2.34(dt,1H,J=4.6,13.3Hz,H-2'a),2.27(s,3H,H-6'),2.22(m,1H,H-2'b).13C NMR(150MHz,CDCl3)δ171.3,165.4(d,1JC-F=254.9Hz),138.8,136.8,135.3,133.5,130.7,129.7×2(d,3JC-F=9.2Hz),127.2,126.6,125.8,125.7,124.1,123.9,123.2,120.9,120.4,118.1,115.9×2(d,2JC-F=22.8Hz),115.8,115.5,112.0,107.7,94.7,86.6,82.3,79.3,60.4,52.1,33.7,29.3,28.2.ESI-MS m/z641.2[M+H]+。
化合物146和147的制备
按照化合物139的合成方法,由131(25.0mg,0.039mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-对氯苯磺酰十字孢碱(146)8.2mg(收率32.0%)和7β-羟基-3′-N-对氯苯磺酰十字孢碱(147)5.9mg(收率23.0%)。
化合物146:1H NMR(600MHz,CDCl3)δ8.63(d,2H,J=7.7Hz,Ar-H),8.23(s,1H,-NH),7.84(m,2H,Ar-H),7.61(m,2H,Ar-H),7.49(d,1H,J=8.4Hz,Ar-H),7.43(t,1H,J=7.3,Ar-H),7.36(t,1H,J=7.7Hz,Ar-H),6.87(t,1H,J=7.7Hz,Ar-H),6.80(d,1H,J=7.7Hz,Ar-H),6.79(dd,1H,J=3.7,11.7Hz,Ar-H),6.54(d,1H,J=7.7Hz,H-7),6.53(m,1H,H-1'),5.43(br s,1H,-OH),4.53(dd,1H,J=6.3,12.4Hz,H-3'),3.60(s,1H,H-4'),2.58(s,3H,4'-OCH3),2.47(s,3H,3'-NCH3),2.34(m,1H,H-2'a),2.22(dt,1H,J=4.2,12.3Hz,H-2'b),1.92(s,3H,H-6').13C NMR(150MHz,CDCl3)δ173.3,139.7,138.3,138.0,135.8,133.7,130.4,129.9×2,128.5×2,126.7,125.9,125.5,124.6,124.4,124.3,122.5,120.7,119.6,117.9,116.3,114.9,111.0,107.1,94.3,86.2,82.2,80.3,59.7,52.2,30.6,29.2,28.7.ESI-MS m/z 657.6/659.6[M+H]+。
化合物147:1H NMR(600MHz,CDCl3)δ9.14(d,1H,J=7.9Hz,Ar-H),8.28(d,1H,J=7.8Hz,-NH),7.76(br d,2H,J=8.0Hz,Ar-H),7.62(d,1H,J=8.5Hz,Ar-H),7.54(br d,2H,J=8.7Hz,Ar-H),7.40(t,1H,J=7.3Hz,Ar-H),7.30(t,1H,J=7.8Hz,Ar-H),7.25(m,2H,Ar-H),7.19(t,1H,J=7.7Hz,Ar-H),7.06(s,1H,H-7),6.92(d,1H,J=8.0Hz,Ar-H),6.41(dd,1H,J=4.5,8.9Hz,H-1'),6.16(s,1H,-OH),4.44(dd,J=5.4,11.9Hz H-3'),3.79(s,1H,H-4'),2.51(s,3H,4'-OCH3),2.43(s,3H,3'-NCH3),2.28(m,1H,J=4.2,13.2Hz,H-2'a),2.19(m,1H,H-2'b),2.15(s,3H,H-6').13C NMR(150MHz,CDCl3)δ171.5,139.7,138.6,137.7,136.7,133.5,130.6,119.9×2,118.4×2,127.0,126.4,135.7,125.6,124.1,123.9,123.1,120.8,120.3,118.0,115.6,115.5,111.8,107.7,94.6,86.6,82.2,79.3,60.2,52.1,30.6,29.2,28.2.ESI-MS m/z 657.6/659.6[M+H]+.
化合物148和149的制备
按照化合物139的合成方法,由132(25.0mg,0.036mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-对溴苯磺酰十字孢碱(148)7.8mg(收率30.5%)和7β-羟基-3′-N-对溴苯磺酰十字孢碱(149)5.5mg(收率21.5%)。
化合物148:1H NMR(600MHz,CDCl3)δ8.90(s,1H,Ar-H),8.66(d,1H,J=7.4Hz,-NH),8.59(d,1H,J=7.8Hz,Ar-H),7.77(m,4H,Ar-H),7.46(d,1H,J=8.1Hz,Ar-H),7.42(d,1H,J=8.5Hz,Ar-H),7.36(t,1H,J=7.7Hz,Ar-H),6.82(dd,1H,J=3.5,9.8Hz,Ar-H),6.78(m,2H,Ar-H),6.56(d,1H,J=10.9Hz,H-7),6.46(t,1H,J=7.2Hz,H-1'),5.78(d,1H,J=11.1Hz,-OH),4.52(dd,1H,J=6.2,12.8Hz,H-3'),3.56(s,1H,H-4'),2.56(s,3H,4'-OCH3),2.48(s,3H,3'-NCH3),2.35(m,1H,H-2'a),2.19(dt,1H,J=2.5,13.5Hz,H-2'b),2.86(s,3H,H-6').13C NMR(150MHz,CDCl3)δ173.2,138.5,138.2,135.6,133.8,132.9×2,130.3,128.6×2,128.1,126.5,125.8,125.4,124.8,124.4,124.2,122.4,120.6,119.5,117.9,116.2,114.8,110.8,107.0,94.2,86.2,82.4,80.5,59.6,52.2,30.5,29.1,28.7.ESI-MS m/z 701.5/703.5[M+H]+。
化合物149:1H NMR(600MHz,CDCl3)δ9.07(d,1H,J=7.8Hz,Ar-H),8.36(d,1H,J=7.8Hz,-NH),7.77(m,4H,Ar-H),7.64(d,1H,J=8.2Hz,Ar-H),7.43(t,1H,J=6.8Hz,Ar-H),7.37(d,1H,J=3.9Hz,Ar-H),7.31(m,2H,Ar-H),7.13(t,1H,J=7.7Hz,Ar-H),7.08(s,1H,H-7),6.99(d,1H,J=8.1Hz,Ar-H),6.51(dd,1H,J=4.0,9.2Hz,H-1'),6.34(s,1H,-OH),4.49(ddd,1H,J=1.9,5.5,12.8Hz,H-3'),3.86(s,1H,H-4'),2.58(s,3H,4'-OCH3),2.48(s,3H,3'-NCH3),2.44(m,1H,H-2'a),2.25(s,3H,H-6'),2.22(m,1H,H-2'b).13C NMR(150MHz,CDCl3)δ171.6,138.7,138.3,136.7,134.4,132.9×2,130.6,128.5×2,128.2,127.1,126.1,126.5,125.7,125.7,124.2,123.1,120.9,120.3,118.0,115.7,115.6,111.8,107.6,94.6,86.6,82.3,79.4,60.3,52.1,33.7,29.3,28.2.ESI-MS m/z 701.5/703.5[M+H]+。
化合物150的制备
按照化合物139的合成方法,由3'-N-对碘苯磺酰十字孢碱(25.0mg,0.034mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-对碘苯磺酰十字孢碱(150)8.1mg(收率31.7%)。1H NMR(600MHz,CDCl3)δ8.65(d,1H,J=7.4Hz,Ar-H),8.60(d,1H,J=7.9Hz,Ar-H),8.57(s,1H,-NH),7.99(m,2H,Ar-H),7.60(m,2H,Ar-H),7.47(d,1H,J=8.0Hz,Ar-H),7.43(t,1H,J=7.3Hz,Ar-H),7.36(t,1H,J=7.3Hz,Ar-H),6.83(t,1H,J=7.8Hz,Ar-H),6.79(t,2H,J=8.0Hz,Ar-H),6.54(d,1H,J=11.8Hz,H-7),6.49(t,1H,J=7.8Hz,H-1'),5.62(d,1H,J=12.2Hz,-OH),4.50(dd,1H,J=5.8,12.8Hz,H-3'),3.57(s,1H,H-4'),2.57(s,3H,4'-OCH3),2.47(s,1H,3'-NCH3),2.34(m,1H,H-2'a),2.20(dt,1H,J=3.2,12.4Hz,H-2'b),1.90(s,3H,H-6').13C NMR(150MHz,CDCl3)δ173.5,139.3×2,138.8×2,138.3,135.7,133.8,130.4,128.5×2,126.6×2,125.8,125.4,124.4,122.4,120.6,119.6,117.9,116.3,114.9,110.9,109.4,100.5,94.3,86.2,82.2,80.4,59.6,52.3,30.6,29.1,28.7.ESI-MS m/z 749.6[M+H]+.
化合物151和152的制备
按照化合物139的合成方法,由133(25.0mg,0.034mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7α-羟基-3′-N-苯磺酰十字孢碱(151)7.9mg(收率30.8%)和7β-羟基-3′-N-苯磺酰十字孢碱(152)5.4mg(收率21.1%)。
化合物151:1H NMR(600MHz,CDCl3)δ8.66(s,1H,-NH),8.63(t,2H,J=7.4Hz,Ar-H),7.91(d,2H,J=7.4Hz,Ar-H),7.70(t,1H,J=7.8Hz,Ar-H),7.64(m,2H,Ar-H),7.40(m,2H,Ar-H),7.34(t,1H,J=7.4Hz,Ar-H),6.77(m,2H,Ar-H),6.73(d,1H,J=7.6Hz,Ar-H),6.54(d,1H,J=10.0Hz,H-7),6.48(d,1H,J=8.0Hz,H-1'),5.75(d,1H,J=10.0Hz,-OH),4.53(dd,1H,J=12.8,5.5Hz,H-3'),3.48(s,1H,H-4'),2.58(s,3H,4'-OCH3),2.42(s,3H,3'-NCH3),2.36(m,1H,H-2'a),2.18(m,1H,H-2'b),1.82(s,3H,H-6').13C NMR(150MHz,CDCl3)δ173.5,139.5,138.2,135.7,133.7,133.2,130.4,129.6×2,127.0×2,126.6,125.7,125.3,124.7,124.3,124.2,122.4,120.5,119.5,117.8,116.1,114.8,110.8,107.0,94.2,85.8,82.2,80.4,59.4,52.1,30.5,29.0,28.6.ESI-MS m/z 623.2[M+H]+.
化合物152:1H NMR(600MHz,CDCl3)δ9.19(d,1H,J=7.8Hz,Ar-H),8.38(d,1H,J=7.4Hz,-NH),7.88(d,2H,J=7.3Hz,Ar-H),7.70(d,1H,J=7.8Hz,Ar-H),7.62(m,3H,Ar-H),7.43(t,1H,J=7.8Hz,Ar-H),7.36(t,1H,J=7.8Hz,Ar-H),7.30(t,1H,J=7.3Hz,Ar-H),7.24(t,1H,J=7.3Hz,Ar-H),6.99(d,1H,J=7.9Hz,ArH),6.83(s,1H,H-7),6.47(dd,1H,J=9.2,4.6Hz,H-1'),6.31(br s,1H,-OH),4.50(ddd,1H,J=1.9,5.3,13.0Hz,H-3'),3.83(s,1H,H-4'),2.61(s,3H,4'-OCH3),2.42(s,3H,3'-NCH3),2.34(m,1H,H-2'a),2.29(s,3H,H-6'),2.22(m,1H,H-2'b).13C NMR(150MHz,CDCl3)δ171.3,139.2,138.8,136.8,133.4,133.2,130.7,129.5×2,127.2,127.1×2,126.6,125.6,125.8,125.7,124.1,124.0,123.3,120.9,120.4,118.1,115.6,112.0,107.7,94.7,86.3,82.4,79.4,60.3,52.1,20.7,29.3,28.1.ESI-MS m/z 623.2[M+H]+。
化合物153的制备
按照化合物130的合成方法,由十字孢碱(46.6mg,0.1mmol)、三乙胺和对氟苯甲酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得3′-N-对氟苯甲酰十字孢碱(153)53.8mg,收率91.5%。1H NMR(600MHz,CDCl3)δ9.45(d,1H,J=8.2Hz,Ar-H),7.87(d,1H,J=7.8Hz,Ar-H),7.78(d,1H,J=7.8Hz,Ar-H),7.48(t,1H,J=7.8Hz,Ar-H),7.46(t,1H,J=7.3Hz,Ar-H),7.43(m,2H,J=7.3Hz,Ar-H),7.37(t,1H,J=7.3Hz,Ar-H),7.33(t,1H,J=7.3Hz,Ar-H),7.21(d,1H,J=7.3Hz,Ar-H),7.10(d,2H,J=7.3Hz,Ar-H),6.94(br s,1H,-NH),6.66(brs,1H,H-1'),5.17(d,1H,J=7.8Hz,H-3'),4.97(d,1H,J=16.0Hz,H-7a),4.93(d,1H,J=16.0Hz,H-7b),4.21(s,1H,H-4'),2.84(s,3H,4'-OCH3),2.72(m,1H,H-2'a),2.65(m,1H,H-2'b),2.53(s,3H,3'-NCH3),2.45(s,3H,H-6').13C NMR(150MHz,CDCl3)δ173.5,171.5,163.6(d,1JC-F=247.2Hz),136.6×2,132.5,132.2,130.6,129.4×2(d,3JC-F=6.9Hz),126.9,126.4,125.5,125.1,124.8,123.7,121.6,120.6,120.2,119.2,116.3,115.8×2(d,2JC-F=13.7Hz),114.6,112.4,107.8,94.7,84.8,82.4,60.5,49.8,46.1,34.6,29.2,28.2.ESI-MS m/z 589.2[M+H]+。
化合物154的制备
按照化合物134的合成方法,由化合物153(15.0mg,0.025mmol)和氯代丁二酰亚胺(7.5mg,0.056mmol)合成。经过凝胶柱色谱分离、甲醇洗脱得3-氯-3′-N-对氟苯甲酰十字孢碱(154)10.5mg,收率33.2%。1H NMR(600MHz,CDCl3)δ9.38(s,1H,Ar-H),7.89(d,1H,J=7.8Hz,Ar-H),7.77(d,1H,J=7.3Hz,Ar-H),7.48(d,2H,J=7.8Hz,Ar-H),7.43(d,2H,J=7.3,Ar-H),7.36(t,1H,J=7.8Hz,Ar-H),7.35(d,1H,J=7.8Hz,Ar-H),7.12(t,2H,J=7.3Hz,Ar-H),7.10(br s,1H,-NH),6.74(br s,1H,H-1'),5.20(d,1H,J=7.8Hz,H-4'),4.98(br s,2H,H-7),4.22(s,1H,H-3'),2.86(s,3H,4'-OCH3),2.75(m,1H,H-2'a),2.62(m,1H,H-2'b),2.57(s,3H,3'-NCH3),2.44(s,3H,H-6').13C NMR(150MHz,CDCl3)δ173.0,171.5,163.6(d,1JC-F=247.2Hz),134.8×2,132.8,132.2,130.4,129.3×2(d,3JC-F=6.9Hz),,127.1,126.1,125.7,125.4,124.7,124.6,121.7,120.8,119.2,115.9×2(d,2JC-F=13.7Hz),115.4,115.0,112.4,108.6×2,94.8,84.7,82.5,60.5,49.8,46.1,34.5,29.2,28.1.ESI-MS m/z 623.3/625.3[M+H]+。
化合物155的制备
按照化合物139的合成方法,由化合物153(20.0mg,0.034mmol)、DMSO和NaOH溶液合成。经过凝胶柱色谱分离、甲醇洗脱得7β-羟基-3′-N-对氟苯甲酰十字孢碱(155)7.6mg,收率37.0%。1H NMR(600MHz,DMSO)δ9.28(d,1H,J=7.7Hz,Ar-H),8.52(d,1H,J=7.7Hz,-NH),7.77(d,1H,J=7.7Hz,Ar-H),7.50(t,1H,J=8.2Hz,Ar-H),7.49(t,1H,J=8.2Hz,Ar-H),7.44(d,2H,J=7.3Hz,Ar-H),7.39(t,1H,J=7.7Hz,Ar-H),7.37(t,1H,J=7.7Hz,Ar-H),7.35(t,1H,J=7.7Hz,Ar-H),7.11(d,2H,J=7.7Hz,Ar-H),7.06(s,1H,H-7),6.71(brs,1H,-OH),6.50(br s,1H,H-1'),5.20(d,1H,J=8.2Hz,H-3'),4.26(s,1H,H-4'),2.87(s,3H,4'-OCH3),2.74(m,1H,H-2'a),2.68(m,1H,H-2'b),2.54(s,3H,3'-NCH3),1.67(s,3H,H-6');13C NMR(150MHz,DMSO)δ171.0,170.9,163.9(d,1JC-F=247.2Hz),137.1×2,135.4,133.5,130.2,129.9×2(d,3JC-F=6.9Hz),126.1×2,125.8,124.0,123.9,123.1,120.7,120.2,119.3×2,116.1,115.7×2(d,2JC-F=13.7Hz),113.9,112.7,109.6,95.2,82.7,82.7,78.9,61.0,49.9,34.6,29.9,27.5;ESI-MS m/z 604.2[M+H]+。
化合物156的制备
按照化合物130的合成方法,由135a(48.0mg,0.1mmol)、三乙胺和对三氟甲基苯甲酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得7-氧亚基-3′-N-对三氟甲基苯甲酰十字孢碱(156)59.5mg,收率91.3%。1H NMR(600MHz,CDCl3)δ9.34(d,1H,J=7.7Hz,Ar-H),9.17(d,1H,J=7.8Hz,Ar-H),7.91(d,2H,J=7.7Hz,Ar-H),7.71(t,1H,J=7.4Hz,Ar-H),7.66(t,1H,J=7.8Hz,Ar-H),7.63(t,1H,J=7.8Hz,Ar-H),7.53(t,2H,J=7.7Hz,Ar-H),7.52(brs,1H,-NH),7.41(t,1H,J=7.8Hz,Ar-H),7.40(d,1H,J=7.8Hz,Ar-H),7.12(d,1H,J=7.4Hz,Ar-H),6.60(dd,1H,J=3.7Hz,9.2Hz,H-1'),4.56(dd,1H,J=5.5Hz,12.4Hz,H-3'),3.83(s,1H,H-4'),2.74(s,3H,4'-OCH3),2.47(m,1H,H-2'a),2.42(s,3H,3'-NCH3),2.38(m,1H,H-2'b),2.26(s,3H,H-6').13C NMR(150MHz,CDCl3)δ169.8,169.6,167.8,139.3,139.2,137.8×2,133.2×2,131.4(q,2JC-F=27.3Hz),130.0,129.6×2,127.1×2,126.5(q,3JC-F=8.4Hz),126.3,123.8(q,1JC-F=270.0Hz),122.5,121.4,121.3,121.0(q,3JC-F=7.8Hz),119.4,117.4,116.4,114.7,111.3,108.2,94.7,86.0,82.5,60.1,51.9,30.7,28.9,28.3;ESI-MS m/z 653.2[M+H]+。
化合物157的制备
按照化合物130的合成方法,由十字孢碱(46.6mg,0.1mmol)、三乙胺和对碘苯磺酰氯合成。经过凝胶柱色谱分离、甲醇洗脱得3′-N-对碘苯磺酰十字孢碱(157)62.1mg,收率84.8%。1H NMR(600MHz,CDCl3)δ9.42(d,1H,J=7.8Hz,Ar-H),7.99(d,2H,J=6.9Hz,Ar-H),7.89(d,1H,J=7.8Hz,Ar-H),7.72(d,1H,J=8.3Hz,Ar-H),7.61(d,2H,J=6.9Hz,Ar-H),7.48(t,1H,J=7.3Hz,Ar-H),7.45(t,1H,J=7.3Hz,Ar-H),7.37(t,1H,J=7.8Hz,Ar-H),7.35(t,1H,J=7.8Hz,Ar-H),7.10(d,1H,J=8.2Hz,Ar-H),6.61(br s,1H,-NH),6.57(dd,1H,J=4.6Hz,9.2Hz,H-1'),4.99(d,1H,J=16.5Hz,H-7a),4.92(d,1H,J=16.5Hz,H-7b),4.50(m,1H,H-4'),3.96(s,1H,H-3'),2.72(s,3H,4'-OCH3),2.47(s,1H,3'-NCH3),2.45(m,1H,H-2'a),2.43(s,3H,H-6'),2.45(m,1H,H-2'a);13C NMR(150MHz,CDCl3)δ173.3,138.9×2,138.5,136.6×2,132.5,130.5,128.4×2,127.0,126.4,125.6,125.3,124.8,123.7,121.7,120.8,120.3,119.3,116.4,114.7,112.3,107.7,100.6,94.7,86.6,82.5,60.5,52.2,46.0,30.9,29.2,28.2.ESI-MS m/z 733.3[M+H]+。
化合物158的制备
i)N-甲基-3,4-二(3-吲哚)马来酰亚胺(158a)的制备
按照化合物82的合成方法,由镁丝(128mg,5.3mmol)、溴代乙烷(396μL,5.3mmol)和吲哚(622mg,5.3mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得红色固体(158a)263mg,收率73%。1H NMR(600MHz,DMSO-d6)δ11.70(s,2H,indole-NH),7.77(d,2H,J=2.3Hz,Ar-H),7.38(d,2H,J=8.2Hz,Ar-H),6.98(t,2H,J=7.7Hz,Ar-H),6.82(d,2H,J=7.8Hz,Ar-H),6.64(t,2H,J=7.8Hz,Ar-H),3.04(s,3H,-CH3).13C NMR(150MHz,DMSO-d6)δ172.4×2,136.6×2,129.7×2,127.6×2,125.9×2,122.2×2,121.5×2,119.9×2,112.3×2,106.2×2,24.5.ESI-MS m/z 342.1[M+H]+。
ii)3,4-二(3-吲哚)马来酸酐(158b)的制备
按照化合物84a的合成方法,由化合物158a(120mg,0.35mmol)和10%的KOH溶液合成,硅胶柱色谱分离、二氯甲烷洗脱得红色固体(158b)104mg,收率90%。1H NMR(600MHz,DMSO-d6)δ11.93(d,2H,J=2.8Hz,indole-NH),7.86(d,2H,J=2.8Hz,Ar-H),7.44(d,2H,J=8.2Hz,Ar-H),7.04(t,2H,J=8.2Hz,Ar-H),6.87(d,2H,J=7.7Hz,Ar-H),6.71(t,2H,J=7.7Hz,Ar-H).13C NMR(150MHz,DMSO-d6)δ167.1×2,136.7×2,131.1×2,125.8×2,125.5×2,122.6×2,121.7×2,120.4×2,112.7×2,105.5×2.ESI-MS m/z 329.1[M+H]+。
iii)3,4-二(3-吲哚)马来酰亚胺(158c)的制备
按照化合物1的制备方法,由化合物158b(100mg,0.3mmol)、HMDS(6.4mL,30.5mmol)、MeOH(0.61mL,15.2mmol)制得橙红色粉末(158c)70mg,收率71%。1H NMR(600MHz,DMSO-d6)δ11.65(brs,2H,indole-NH),10.89(brs,1H,imide-NH),7.72(d,2H,J=2.8Hz,Ar-H),7.36(d,2H,J=8.2Hz,Ar-H),6.96(dt,2H,J=8.2Hz,1.0Hz,Ar-H),6.79(d,2H,J=7.8Hz,Ar-H),6.61(dt,2H,J=8.2Hz,1.0Hz,Ar-H).13C NMR(150MHz,DMSO-d6)δ173.6×2,136.4×2,129.6×2,128.2×2,125.9×2,122.1×2,121.4×2,119.8×2,112.3×2,106.1×2.ESI-MS m/z 328.2[M+H]+。
ⅳ)N-羟甲基-3,4-二(1-羟甲基-3-吲哚)马来酰亚胺(158)的制备
按照化合物4的制备方法,从158c(76.5mg,0.23mmol)、NaHCO3(98mg,1.17mmol)和甲醛溶液(3mL,质量分数37%)制得红色粉末状固体(158)96mg,收率99%。1H NMR(600MHz,DMSO-d6)δ7.95(s,2H,Ar-H),7.55(d,2H,J=8.2Hz,Ar-H),7.03(t,2H,J=7.3Hz,Ar-H),6.76(d,2H,J=8.2Hz,Ar-H),6.64(t,2H,J=7.3Hz,Ar-H),5.60(s,4H,indole-CH2 -OH),4.98(s,2H,imide-CH2 -OH).13C NMR(150MHz,DMSO-d6)δ171.6×2,136.0×2,132.4×2,127.6×2,127.0×2,122.4×2,121.5×2,120.6×2,111.4×2,105.9×2,69.7,60.9×2.HR-ESIMS m/z 440.1236[M+H]+(calcd.for C23H19N3O5Na,440.1222).
化合物159的制备
按照化合物14的制备方法,由化合物8b(30mg,0.055mmol)和乙二胺(0.5mL)反应,硅胶柱色谱分离、二氯甲烷:甲醇=8:1(v/v)洗脱得深红色固体N-(2-氨乙基)-2,3-二(1-氰丙基-3-吲哚)马来酰亚胺(159)30mg,收率94%。1H NMR(600MHz,DMSO-d6)δ7.76(s,2H,Ar-H),7.50(d,2H,J=8.2Hz,Ar-H),7.07(t,2H,J=7.6Hz,Ar-H),6.89(d,2H,J=7.9Hz,Ar-H),6.72(s,2H,Ar-H),4.28(t,4H,J=5.5Hz,-N-CH2 -(CH2)2CN),3.63(t,2H,J=5.7Hz,-N-CH2 -CH2NH2),2.86(t,2H,J=5.7Hz,-NCH2-CH2 -NH2),2.37(t,J=6.8Hz,4H,N(CH2)2-CH2 -CN),2.03-1.98(m,4H,N-CH2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ171.9×2,136.3×2,132.4×2,127.4×2,126.0×2,122.5×2,121.9×2,120.3×2,120.1×2,110.6×2,105.7×2,54.0,45.0×2,39.9,26.0×2,14.2×2.ESI-MS m/z 505.1[M+H]+.
化合物160的制备
按照化合物24的制备方法,由化合物8b(60mg,0.129mmol)、4-(2-氨乙基)吗啉(120μL,0.909mmol)和催化量Et3N合成,硅胶柱色谱分离、二氯甲烷:甲醇=30:1(v/v)洗脱得红色固体N-(2-(4-吗啉)乙基)-2,3-二(1-氰丙基-3-吲哚)马来酰亚胺(160)44mg,收率59%。1H NMR(500MHz,DMSO-d6)δ7.77(s,2H,Ar-H),7.51(d,2H,J=8.3Hz,Ar-H),7.07(t,2H,J=7.6Hz,Ar-H),6.87(d,2H,J=8.0Hz,Ar-H),6.73(t,2H,J=7.5Hz,Ar-H),4.27(t,4H,J=6.6Hz,-N-CH2 -(CH2)2CN),3.69(t,2H,J=6.0Hz,-N-CH2 -CH2-morpholine),3.52(t,4H,J=4.5Hz,morpholine-N(CH2CH 2)2O),2.53(t,2H,J=6.0Hz,-NCH2-CH2 -morpholine),2.42(t,4H,J=5.3Hz,N(CH2)2-CH2 -CN),2.37(t,4H,J=3.9Hz,morpholine-N(CH 2CH2)2O),2.03-1.98(m,4H,NCH2-CH2 -CH2CN).13C NMR(125MHz,DMSO-d6)δ171.8×2,136.3×2,132.5×2,127.2×2,126.0×2,122.5×2,121.8×2,120.4×2,120.2×2,110.6×2,105.6×2,66.7×2,56.4,53.6×2,45.0×2,35.3,26.0×2,14.2×2.ESI-MS m/z 575.2[M+H]+.
化合物161的制备
按照化合物24的合成方法,由化合物8b(60mg,0.129mmol)、4-(2-氨乙基)哌嗪(100μL,0.90mmol)和催化量Et3N合成,硅胶柱色谱分离、二氯甲烷:甲醇=10:1(v/v)洗脱得红色固体N-(2-哌嗪乙基)-2,3-二(1-氰丙基-3-吲哚)马来酰亚胺(161)44mg,收率75%。1HNMR(600MHz,DMSO-d6)δ7.77(s,2H,Ar-H),7.51(d,2H,J=8.3Hz,Ar-H),7.07(t,2H,J=7.6Hz,Ar-H),6.87(d,2H,J=8.0Hz,Ar-H),6.73(t,2H,J=7.5Hz,Ar-H),4.27(t,4H,J=6.5Hz,-N-CH2 -(CH2)2CN),3.67(t,4H,J=6.1Hz,piperazine-N(CH2CH2)2NH),2.73(t,4H,J=4.9Hz,piperazine-N(CH2CH 2)2NH),2.52(t,2H,J=6.1Hz,imide-NCH 2-CH2-piperazine),2.43(t,2H,J=4.9Hz,imide-NCH2-CH 2-piperazine),2.37(t,4H,J=7.1Hz,N(CH2)2-CH2 -CN),1.99-2.02(m,4H,NCH2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ171.8×2,136.3×2,132.5×2,127.3×2,126.0×2,122.5×2,121.8×2,120.3×2,120.2×2,110.6×2,105.6×2,56.4,53.2×2,45.3×2,45.0×2,35.4,26.0×2,14.2×2.ESI-MS m/z 574.2[M+H]+.
化合物162的制备
以6ml干燥吡啶溶解化合物8b(55mg,0.119mmol),加入盐酸羟胺(17mg,0.238mmol),氮气保护下100℃下反应1.5h。TLC检测至反应不再进行,冷却至室温,旋蒸至干。硅胶柱色谱分离、二氯甲烷:甲醇=15:1(v/v)洗脱得红色固体N-羟基-2,3-二(1-氰丙基-3-吲哚)马来酰亚胺(162)51mg,收率89%。1H NMR(600MHz,DMSO-d6)δ10.46(s,1H,-OH),7.78(s,2H,Ar-H),7.51(d,2H,J=8.3Hz,Ar-H),7.07(t,2H,J=7.6Hz,Ar-H),6.86(d,2H,J=8.0Hz,Ar-H),6.72(t,2H,J=7.5Hz,Ar-H),4.28(t,4H,J=6.5Hz,-N-CH2 -(CH2)2CN),2.37(t,4H,J=7.1Hz,N(CH2)2-CH2 -CN),2.04-1.98(m,4H,NCH2-CH2 -CH2CN).13C NMR(150MHz,DMSO-d6)δ168.6×2,136.3×2,132.6×2,125.9×2,124.7×2,122.5×2,121.8×2,120.4×2,120.3×2,110.7×2,105.5×2,45.0×2,26.0×2,14.2×2.ESI-MS m/z478.2[M+H]+.
化合物163的制备
按照化合物162的合成方法,由化合物26e(55mg,0.13mmol)和盐酸羟胺(25mg,0.25mmol)合成,硅胶柱色谱分离、二氯甲烷:甲醇=20:1(v/v)洗脱得红色固体N-羟基-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(162)58mg,收率99%。1H NMR(600MHz,DMSO-d6)δ11.83(s,1H,indole-NH),10.50(brs,1H,-OH),7.84(s,1H,Ar-H),7.79(s,1H,Ar-H),7.57(s,1H,Ar-H),7.47(d,1H,J=8.2Hz,Ar-H),7.03(t,1H,J=7.6Hz,Ar-H),6.76(t,1H,J=7.6Hz,Ar-H),6.74(d,1H,J=7.1Hz,Ar-H),6.68(t,1H,J=6.2Hz,Ar-H),6.67(d,1H,J=8.1Hz,Ar-H),4.25(q,2H,J=7.0Hz,-CH2 -CH3),1.32(t,3H,J=7.1Hz,-CH2-CH3 ).13C NMR(150MHz,DMSO-d6)δ168.7×2,137.3,136.0,132.3,130.7,126.1,125.2,124.7,124.3,122.9,122.7,122.3,121.6,120.2,114.9×2,110.7,106.1,105.0,41.2,15.7.HR-ESIMS m/z 450.0459[M+H]+(calcd.for C22H16N3O3Br,449.0375).
化合物164的制备
按照化合物24的制备方法,由化合物26e(30mg,0.069mmol)、4-(2-氨乙基)吗啉(64μL,0.484mmol)和催化量Et3N合成,硅胶柱色谱分离、二氯甲烷:甲醇=25:1(v/v)洗脱得红色固体N-(2-(4-吗啉)乙基)-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(164)33mg,收率88%。1H NMR(500MHz,DMSO-d6)δ11.78(s,1H,indole-NH),7.81(s,1H,Ar-H),7.76(s,1H,Ar-H),7.56(d,1H,J=1.5Hz,Ar-H),7.46(d,1H,J=8.3Hz,Ar-H),7.03(t,1H,J=7.6Hz,Ar-H),6.78(d,1H,J=7.7Hz,Ar-H),6.74(dd,1H,J=8.4Hz,1.6Hz,Ar-H),6.69(d,1H,J=8.4Hz,Ar-H),6.67(t,1H,J=8.2Hz,Ar-H),4.24(q,2H,J=7.2Hz,-CH2 -CH3),3.67(t,2H,J=6.5Hz,imide-NCH2 CH2-),3.51(t,4H,J=4.2Hz,morpholine-N(CH2-CH 2)2O),2.52(t,2H,J=6.5Hz,imide-NCH2CH2 -),2.40(t,4H,J=4.2Hz,morpholine-N(CH 2-CH2)2O),1.32(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.9,171.8,137.3,136.0,132.1,130.5,127.7,126.4,126.2,124.7,122.9,122.6,122.3,121.6,120.1,114.9,114.8,110.7,106.2,105.1,66.6×2,56.3,53.6×2,41.2,35.3,15.7.HR-ESIMS m/z547.1354[M+H]+(calcd.for C28H27N4O3Br,546.1267).
化合物165的制备
按照化合物24的合成方法,由化合物26e(30mg,0.069mmol)、4-(2-氨乙基)哌嗪(93μL,0.712mmol)和催化量Et3N合成,硅胶柱色谱分离、二氯甲烷:甲醇=8:1(v/v)洗脱得红色固体N-(2-哌嗪乙基)-2-(1-乙基-3-吲哚)-3-(6-溴-3-吲哚)马来酰亚胺(165)55mg,收率99%。1H NMR(500MHz,DMSO-d6)δ11.78(s,1H,indole-NH),7.81(s,1H,Ar-H),7.76(d,1H,J=2.6Hz,Ar-H),7.57(d,1H,J=1.5Hz,Ar-H),7.48(d,1H,J=8.3Hz,Ar-H),7.04(t,1H,J=7.6Hz,Ar-H),6.78(d,1H,J=8.0Hz,Ar-H),6.76(dd,1H,J=8.4Hz,1.6Hz,Ar-H),6.69(d,1H,J=8.5Hz,Ar-H),6.68(t,1H,J=7.8Hz,Ar-H),4.25(q,2H,J=7.1Hz,-CH2 -CH3),3.67(d,2H,J=6.2Hz,imide-NCH2 CH2-),3.15(brs,1H,piperazine-NH),3.03(t,4H,J=4.9Hz,piperazine-N(CH 2-CH2)2NH),2.64(t,4H,J=4.9Hz,piperazine-N(CH2-CH 2)2NH),2.60(t,2H,J=5.8Hz,imide-NCH2CH2 -),1.32(t,3H,J=7.2Hz,-CH2-CH3 ).13C NMR(125MHz,DMSO-d6)δ171.9,171.8,137.3,136.0,132.2,130.6,127.7,126.4,126.1,124.7,122.9,122.6,122.3,121.6,120.2,114.9,114.8,110.8,106.2,105.1,55.5×2,49.6,43.5×2,41.2,35.3,15.7.HR-ESIMS m/z 546.1515[M+H]+(calcd.for C28H28N5O2Br,545.1426).
化合物166的制备
按照化合物64的合成方法,由化合物8(50.0mg,0.108mmol)和I2(2.0mg,0.008mmol)合成,硅胶柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得黄色粉末12,13-二氰丙基-6H-吲哚[2,3-a]吡咯[3,4-c]咔唑-5,7-二酮(166)23mg,收率45%。1H NMR(600MHz,DMSO-d6)δ11.2(s,1H,-NH),9.14(d,2H,J=7.8Hz,Ar-H),7.89(d,2H,J=8.2Hz,Ar-H),7.66(t,2H,J=7.8Hz,Ar-H),7.44(t,2H,J=7.8Hz,Ar-H),4.81(t,4H,J=6.9Hz,N-CH 2-(CH2)2CN),2.13(t,4H,J=6.9Hz,N(CH2)2-CH 2-CN),1.71(m,4H,NCH2-CH 2-CH2CN).13CNMR(150MHz,DMSO-d6)δ171.2×2,144.3×2,133.2×2,128.1×2,125.5×2,123.9×2,122.2×2,121.6×2,120.5×2,120.2×2,113.3×2,47.5×2,24.2×2,14.2×2.HR-ESIMSm/z 458.1604[M–H]–(calcd.for C28H20N5O2,458.1617).
化合物167的制备
0℃下,以THF(5mL)溶解十字孢碱(60mg,0.129mmol),加入二异丙基乙胺(64μL,0.38mmol)和三光气(19mg,0.064mmol),室温搅拌2h,倒入冰水中,乙酸乙酯萃取(2次×30mL),卤水洗(2次×30mL),以无水硫酸钠干燥有机相,旋蒸至干。将粗产物溶于THF(3mL),二异丙基乙胺(127μL,0.77mmol),咪唑(18mg,0.25mmol)和对二甲氨基吡啶(31.5mg,0.258mmol),60℃搅拌反应2h,倒入冰水中,乙酸乙酯萃取(2次×30mL),卤水洗(2次×30mL),以无水硫酸钠干燥有机相,旋蒸至干。硅胶柱色谱分离、二氯甲烷:甲醇=50:1(v/v)洗脱得到淡黄色固体:3′-N-(1-咪唑甲酰)十字孢碱(167)56mg,收率80%。[α]D 18+176(c0.07,CHCl3);1H NMR(500MHz,DMSO-d6)δ9.31(d,J=7.9Hz,1H,ArH),8.68(s,1H,ArH),8.61(s,1H,NH),8.07(d,J=7.8Hz,1H,ArH),8.01(d,J=8.5Hz,1H,ArH),7.79(brs,1H,ArH),7.59(d,J=8.2Hz,1H,ArH),7.51(t,J=8.2Hz,1H,ArH),7.49(t,J=8.2Hz,1H,ArH),7.37(t,J=7.7Hz,1H,ArH),7.35(brs,1H,ArH),7.31(t,J=7.5Hz,1H,ArH),7.04(dd,J=8.5,5.6Hz,1H,H-1'),5.00(s,2H,H-7),4.68(m,1H,H-3'),4.50(brs,1H,H-4'),2.94-2.99(m,1H,H-2'a),2.89(s,3H,3'-NCH3),2.61(s,3H,6'-CH3),2.36-2.43(m,1H,H-2'b),2.39(s,3H,6'-CH3).13C NMR(125MHz,DMSO-d6)δ172.0,150.8,138.6,137.2,136.3,132.6,129.5,125.9,125.8,125.5,125.5,125.3,123.9,122.8,121.7,120.6,119.7,119.6,119.5,115.2,114.3,113.2,109.0,94.7,83.2,82.1,60.2,52.3,45.5,33.2,28.9,27.0.HRESI-MSm/z 561.2242[M+H]+(calcd for C32H29N6O4,561.2245).
化合物168的制备
将十字孢碱(186mg,0.4mmol)用10mL二氯甲烷溶解,室温下加入1mL三乙胺和N,N′-硫羰基二咪唑(214mg,1.2mmol),室温过夜反应。反应液倒入20mL冰水中,二氯甲烷萃取,无水硫酸钠干燥有机相后浓缩,硅胶柱色谱分离、二氯甲烷:甲醇=20:1(v/v)洗脱得到3′-N-(1-咪唑硫代甲酰)十字孢碱(168)180mg,产率78%。1H NMR(500MHz,DMSO-d6)δ9.32(d,J=7.9Hz,1H),8.63(s,1H,NH),8.12(s,1H,ArH),8.07(d,J=7.7Hz,1H,ArH),8.03(d,J=8.5Hz,1H,ArH),7.64(d,J=7.2Hz,1H,ArH),7.59(brs,1H,ArH),7.51(t,J=7.4Hz,1H,ArH),7.50(t,J=7.7,1H,ArH),7.37(t,J=7.4Hz,1H,ArH),7.32(t,J=7.5Hz,1H,ArH),7.13(brs,1H,ArH),7.06(brs,1H,H-1'),5.47(brs,1H,H-3'),5.01(s,2H,H-7),4.79(brs,1H,H-4'),3.06(s,3H,3'-NCH3),3.02-3.09(m,1H,H-2'a),2.73(s,3H,4'-OCH3),2.44(s,3H,6'-CH3),2.41-2.47(m,1H,H-2'b);13C NMR(125MHz,DMSO-d6)δ179.3,171.9,138.8,137.7,136.2,132.7,129.3,129.0,125.8,125.4,125.2×2,123.9,122.7,121.6,120.5,119.9,119.6,119.6,115.3,114.3,113.5,108.9,94.8,81.9×2,60.4,58.1,45.5,38.2,29.4,27.1;HRESI-MS m/z 577.2031[M+H]+(calcd for C32H29N6O3S,577.2022).
化合物169的制备
将化合物168(80mg,0.14mmol)溶于10mL乙腈中,加入碘甲烷(86μL,1.39mmol),室温反应24小时。反应液直接浓缩,用50mL石油醚:二氯甲烷=4:1(v/v)的混合溶液洗纯即得化合物168的咪唑部分的碘甲烷盐75mg,产率76%;HRESI-MS m/z 591.2164[M-I]+(calcdfor C33H31N6O3S,591.2173)。将色胺(2.0g,12.5mmol)溶于20mL四氢呋喃中,降至10℃,依次加入三乙胺(3.5mL,25mmol)和叔丁氧甲酸酐(3.03g,15.0mmol),10℃反应1小时,将反应液倒入100mL冰水中,乙酸乙酯萃取,无水硫酸钠干燥有机相后浓缩,快速柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得到N-[2-(3-吲哚)乙基]氨基甲酸叔丁酯3.16g,产率97%;ESI-MS m/z 261.3[M+H]+。将N-[2-(3-吲哚)乙基]氨基甲酸叔丁酯(1.8g,6.92mmol)溶于110mL THF/H2O(10:1),降至0℃,加入DDQ(3.1g,13.8mmol)并在此温度下反应两小时。反应液倒入200mL乙酸乙酯中,用饱和碳酸氢钠溶液洗至无色,乙酸乙酯层用无水硫酸钠干燥后浓缩,快速柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得到N-[2-氧亚基-2-(3-吲哚)乙基]氨基甲酸叔丁酯1.2g,产率63%;ESI-MS m/z 275.4[M+H]+。将N-[2-氧亚基-2-(3-吲哚)乙基]氨基甲酸叔丁酯(200mg,0.73mmol)用5mL三氟乙酸溶解,10℃下反应一小时,加入苯(5mL×3次)共沸除去三氟乙酸即得到N-[2-氧亚基-2-(3-吲哚)乙基]三氟乙酸铵185mg,产率93%。将化合物168的咪唑部分的碘甲烷盐(75.0mg,0.105mmol)溶于2mL DMF中,加入三乙胺(73.0μL,0.525mmol)和N-[2-氧亚基-2-(3-吲哚)乙基]三氟乙酸铵(85.4mg,0.315mmol,3.0equiv),室温反应24小时。反应液用10mL乙酸乙酯稀释,用1N的盐酸洗,无水硫酸钠干燥后浓缩。半制备HPLC分离、MeOH:H2O=9:1(v/v)洗脱得到3′-N-[N-(2-氧亚基-2-(3-吲哚)乙基)氨基硫代甲酰]十字孢碱(169)40mg,产率56%。[α]D 18+248(c 0.07,CHCl3);1H NMR(500MHz,DMSO-d6)δ12.02(d,J=2.0Hz,1H,NH),9.30(d,J=8.0Hz,1H,ArH),8.59(s,1H,NH),8.49(d,J=2.9Hz,1H,ArH),8.19(d,J=7.0Hz,1H,ArH),8.05(d,J=7.8Hz,1H,ArH),7.98(d,J=8.5Hz,1H,ArH),7.92(t,J=6.0Hz,1H,NH),7.72(d,J=8.3Hz,1H,ArH),7.52(d,J=7.2Hz,1H,ArH),7.49(t,J=7.5Hz,2H,ArH),7.36(t,J=7.4Hz,1H,ArH),7.31(t,J=7.5Hz,1H,ArH),7.25(dt,J=7.7,1.6Hz,1H,ArH),7.21(dt,J=7.6,1.5Hz,1H,ArH),7.07(t,J=7.4Hz,1H,H-1'),5.94(d,J=12.2Hz,1H,H-3'),4.97-5.09(m,2H,H-3”),5.01(s,2H,H-7),4.51(brs,1H,H-4'),2.96(s,3H,3'-NCH3),2.84(s,3H,4'-OCH3),2.72-2.76(m,1H,H-2'a),2.36(s,3H,6'-CH3),2.31(ddd,J=12.5,12.5,6.5Hz 1H,H-2'b);13C NMR(125MHz,DMSO-d6)δ190.2,182.5,172.1,139.1,136.5,136.4,133.4,132.8,129.2,125.7,125.5×2,125.1,125.1,123.9,123.0,122.7,121.9,121.5,121.3,120.4,119.6,119.4,115.4,114.3×2,113.9,112.3,109.2,95.1,83.1,82.5,60.5,54.3,52.0,45.6,32.8,29.6,27.8;HRESI-MS m/z 683.2462[M+H]+(calcd for C39H35N6O4S,683.2441).
化合物170的制备
将溴代苯乙酮(1.97g,10.0mmol)用40mL氯仿溶解,加入六次甲基四胺(1.47g,10.5mmol),室温反应4小时。反应液过滤后将滤渣用80mL甲醇溶解,加入4mL浓盐酸,回流反应3小时。反应液浓缩后,甲醇重结晶得到化合物N-(2-氧亚基-2-苯乙基)氯化铵1.5g,产率88%;ESI-MS m/z 136.3[M-Cl]+。将化合物168的咪唑部分的碘甲烷盐(49.0mg,0.068mmol)溶于2mL DMF中,加入三乙胺(47.2μL,0.34mmol)和N-(2-氧亚基-2-苯乙基)氯化铵(34.9mg,0.204mmol),室温反应24小时。反应液用10mL乙酸乙酯稀释,用1N的盐酸洗,无水硫酸钠干燥有机相后浓缩。快速柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得到3′-N-[N-(2-氧亚基-2-苯乙基)氨基硫代甲酰]十字孢碱(170)18.0mg,产率41%。[α]D 18+257(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.29(d,J=8.1Hz,1H,ArH),8.61(s,1H,NH),8.06(d,J=7.8Hz,1H,ArH),8.04(d,J=7.7Hz,2H,ArH),8.00(d,J=8.5Hz,1H,ArH),7.74(d,J=8.2Hz,1H,ArH),7.68(t,J=7.3Hz,1H,ArH),7.58(t,J=7.6Hz,2H,ArH),7.49(t,J=7.6Hz,2H,ArH),7.36(t,J=7.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),7.08(t,J=7.7Hz,1H,H-1'),5.85(m,1H,H-3'),5.02-5.15(m,2H,H-3”),5.01(s,2H,H-7),4.51(brs,1H,H-4'),2.93(s,3H,3'-NCH3),2.85(s,3H,4'-OCH3),2.70-2.74(m,1H,H-2'a),2.34(s,3H,6'-CH3),2.28(ddd,J=13.0,13.0,7.2Hz,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ195.4,182.5,171.9,139.1,136.3,135.5,133.4,132.8,129.0,128.8×2,127.8×2,125.7,125.4,125.0,124.9,123.8,122.6,121.4,120.3,119.5,119.4,115.3,114.2,114.0,109.1,94.9,82.7,82.3,60.4,54.4,51.9,45.5,32.9,29.6,27.6;HRESI-MS m/z644.2351[M+H]+(calcd for C37H34N5O4S,644.2332).
化合物171的制备
将化合物167(50mg,0.089mmol)溶于10mL乙腈中,加入碘甲烷(55μL,0.89mmol),室温反应24小时。反应液直接浓缩,用50mL石油醚:二氯甲烷=4:1(v/v)的混合溶液洗纯即得化合物167的咪唑盐48mg,产率77%;HRESI-MS m/z 575.2393[M–I]+(calcd forC33H31N6O4,575.2401)。将化合物167的咪唑部分的碘甲烷盐(48.0mg,0.068mmol)溶于2mLDMF中,加入三乙胺(47.2μL,0.34mmol)和N-[2-氧亚基-2-(3-吲哚)乙基]三氟乙酸铵(55.2mg,0.204mmol),室温反应24小时。反应液用10mL乙酸乙酯稀释,用1N的盐酸洗,无水硫酸钠干燥有机相后浓缩。快速柱色谱分离、二氯甲烷:乙酸乙酯=1:3(v/v)洗脱得到3′-N-[N-(2-氧亚基-2-(3-吲哚)乙基)氨基甲酰]十字孢碱(171)36.0mg,产率79%。[α]D 18+105(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ12.08(d,J=2.5Hz,1H,NH),9.29(d,J=8.0Hz,1H,ArH),8.61(s,1H,NH),8.49(d,J=3.1Hz,1H,ArH),8.21(d,J=7.7Hz,1H,ArH),8.04(d,J=7.9Hz,1H,ArH),7.96(d,J=8.4Hz,1H,ArH),7.70(d,J=8.4Hz,2H,ArH),7.52(d,J=8.0Hz,1H,ArH),7.48(t,J=7.6Hz,2H,ArH),7.35(t,J=7.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),7.24(t,J=7.3Hz,1H,ArH),7.21(t,J=7.6Hz,1H,ArH),7.02(t,J=7.6Hz,1H,H-1'),6.84(t,J=5.3Hz,1H,NH),5.00(s,2H,H-7),4.85(d,J=12.5Hz,1H,H-3'),4.50(d,J=5.7Hz,2H,H-3”),4.24(brs,1H,H-4'),2.84(s,3H,3'-NCH3),2.75(s,3H,4'-OCH3),2.61-2.65(m,1H,H-2'a),2.30(s,3H,6'-CH3),2.22(ddd,J=12.5,12.5,7.0Hz,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ192.0,172.2,158.4,139.2,136.6,136.5,133.5,132.9,129.4,125.9,125.7,125.6,125.2×2,124.0,123.1,122.8,122.0,121.6,121.4,120.5,119.7,119.5,115.4,114.3×2,114.0,112.4,109.3,95.1,84.1,82.7,60.6,49.1,47.6,45.7,30.0,29.7,27.6;HRESI-MS m/z 667.2661[M+H]+(calcd for C39H35N6O5,667.2669).
化合物172的制备
将化合物167的咪唑部分的碘甲烷盐(48.0mg,0.068mmol)溶于2mL DMF中,加入三乙胺(47.2μL,0.34mmol)和N-(2-氧亚基-2-苯乙基)氯化铵(34.9mg,0.204mmol),室温反应24小时。反应液用10mL乙酸乙酯稀释,用1N的盐酸洗,无水硫酸钠干燥有机相后浓缩。快速柱色谱分离、二氯甲烷:乙酸乙酯=2:1(v/v)洗脱得到3′-N-[N-(2-氧亚基-2-苯乙基)氨基甲酰]十字孢碱(172)20.0mg,产率47.0%。[α]D 18+174(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.28(d,J=7.9Hz,1H,ArH),8.60(s,1H,NH),8.04(t,J=8.2Hz,1H,ArH),8.03(d,J=8.1Hz,1H,ArH),8.03(d,J=8.3Hz,1H,ArH),7.96(t,J=8.7Hz,1H,ArH),7.70(d,J=8.5Hz,1H,ArH),7.68(t,J=7.6Hz,1H,ArH),7.58(d,J=7.6Hz,1H,ArH),7.57(d,J=7.6Hz,1H,ArH),7.49(t,J=6.8Hz,2H,ArH),7.35(t,J=7.5Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),7.02(t,J=7.6Hz,1H,H-1'),6.90(t,J=5.3Hz,1H,NH),5.00(s,2H,H-7),4.81(d,J=12.5Hz,1H,H-3'),4.59(d,J=5.6Hz,2H,H-3”),4.20(brs,1H,H-4'),2.83(s,3H,3'-NCH3),2.72(s,3H,4'-OCH3),2.59-2.63(m,1H,H-2'a),2.29(s,3H,6'-CH3),2.20(ddd,J=12.5,12.5,6.9Hz,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ196.9,172.2,158.2,139.2,136.5,135.4,133.6,132.9,129.3,129.0×2,128.0×2,125.8,125.6,125.1×2,123.9,122.8,121.6,120.5,119.7,119.5,115.4,114.3,114.0,109.2,95.0,84.0,82.6,60.6,49.1,47.7,45.6,30.0,29.7,27.5;HRESI-MS m/z 628.2568[M+H]+(calcd forC37H34N5O5,628.2560).
化合物173的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(15.0mg,0.021mmol)与DMF、Et3N和4-(2-哌嗪乙基)吗啉合成。半制备HPLC分离、MeOH:H2O=4:1(v/v)洗脱得到3′-N-[4-(2-(4-吗啉)乙基)哌嗪硫代甲酰]十字孢碱(173)8.0mg,产率47.1%。[α]D 20+101°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.29(d,J=7.9Hz,1H,ArH),8.52(s,1H,NH),8.03(d,J=7.8Hz,1H,ArH),7.94(d,J=8.6Hz,1H,ArH),7.56(d,J=8.2Hz,1H,ArH),7.48(t,J=7.8Hz,1H,ArH),7.45(t,J=7.8Hz,1H,ArH),7.34(t,J=7.4Hz,1H,ArH),7.27(t,J=7.5Hz,1H,ArH),7.00(dd,J=8.7,5.5Hz,1H,H-1'),5.23-5.27(m,1H,H-3'),4.98(s,2H,H-7),4.68(brs,1H,H-4'),3.91(brs,4H,morpholine-N(CH2-CH2 )2O),3.50-3.75(m,16H,-N(CH2 -CH2 )2N,piperazine-CH2 -CH2 -morpholine,-N(CH2 -CH2)2O),3.01(s,3H,3'-NCH3),2.94-2.98(m,1H,H-2'a),2.56(s,3H,4'-OCH3),2.44(s,3H,6'-CH3),2.37-2.41(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ193.5,172.0,138.5,136.2,132.4,129.6,125.8,125.5,125.2,125.1,123.9,122.7,121.4,120.4,119.5,119.4,115.2,114.2,113.1,108.8,94.9,83.0,82.2,63.3×2,59.9,56.3,51.6×2,51.0×2,49.8×2,49.2,47.6,45.5,37.3,29.2,27.6;ESI-MS m/z 708.4[M+H]+.
化合物174的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(18.0mg,0.021mmol)与DMF、Et3N和2,6-二氟苯甲胺合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=4:1(v/v)洗脱得到3′-N-[N-(2,6-二氟苯甲基)氨基硫代甲酰]十字孢碱(174)8.0mg,产率48.0%。[α]D 20+128°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.27(d,J=7.9Hz,1H,ArH),8.60(s,1H,NH),8.04(d,J=7.8Hz,1H,ArH),7.99(d,J=8.5Hz,1H,ArH),7.93(t,J=4.2Hz,1H,NH),7.70(d,J=8.3Hz,1H,ArH),7.47(t,J=7.6Hz,2H,ArH),7.39(m,1H,ArH),7.35(t,J=7.5Hz,1H,ArH),7.29(t,J=7.5Hz,1H,ArH),7.06(t,J=7.7Hz,2H,ArH),7.03-7.06(m,1H,H-1'),5.90(d,J=12.5Hz,1H,H-3'),4.99(s,2H,H-7),4.75-4.88(m,2H,H-3”),4.47(brs,1H,H-4'),2.82(s,3H,3'-NCH3),2.74(s,3H,4'-OCH3),2.66-2.71(m,1H,H-2'a),2.35(s,3H,6'-CH3),2.20-2.27(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ181.6,172.0,161.3×2(dd,1JC-F=247.7Hz,3JC-F=8.2Hz),139.0,136.4,132.8,129.6(t,3JC-F=10.5Hz),129.2,125.7,125.4,125.1,125.0,123.8,122.7,121.5,120.4,119.6,119.4,115.3,114.3(t,2JC-F=18.1Hz),114.2,113.9,111.5×2(d,2JC-F=20.1Hz),109.2,95.0,83.0,82.4,60.3,54.2,45.5,38.3,32.9,29.5,27.7;ESI-MS m/z 652.3[M+H]+.
化合物175的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(18.0mg,0.021mmol)与DMF、Et3N和3-氯-4-氟苯甲胺合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=4:1(v/v)洗脱得到3′-N-[N-(3-氯-4-氟苯甲基)氨基硫代甲酰]十字孢碱(175)8.0mg,产率48.0%。[α]D 20+166°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.28(d,J=7.9Hz,1H,ArH),8.62(s,1H,NH),8.26(t,J=4.5Hz,1H,NH),8.05(d,J=7.8Hz,1H,ArH),8.00(d,J=8.5Hz,1H,ArH),7.71(d,J=8.2Hz,1H,ArH),7.51(d,J=7.1Hz,1H,ArH),7.48(t,J=7.7Hz,2H,ArH),7.38(t,J=7.8Hz,1H,ArH),7.32-7.37(m,2H,ArH),7.29(t,J=7.5Hz,1H,ArH),7.07(t,J=7.6Hz,1H,H-1'),5.92(d,J=12.9Hz,1H,H-3'),5.00(s,2H,H-7),4.79-4.87(m,2H,H-3”),4.50(brs,1H,H-4'),2.88(s,3H,3'-NCH3),2.79(s,3H,4'-OCH3),2.68-2.72(m,1H,H-2'a),2.36(s,3H,6'-CH3),2.26(ddd,J=12.9,12.9,6.9Hz,1H,H-2'b);13CNMR(150MHz,DMSO-d6)δ182.1,172.0,156.1(d,1JC-F=244.8Hz),139.1,137.8,136.4,132.8,129.2,129.1(d,3JC-F=7.4Hz),127.9(d,3JC-F=7.4Hz),125.7,125.4,125.1,125.0,123.8,122.7,121.5,120.4,119.6,119.4,119.0(d,2JC-F=17.8Hz),116.6(d,2JC-F=20.9Hz),115.3,114.2,113.9,109.2,95.0,83.0,82.4,60.4,54.4,45.5,35.9,33.0,29.6,27.7;ESI-MS m/z 668.4/670.4[M+H]+.
化合物176的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(18.0mg,0.021mmol)与DMF、Et3N和2-氯-6-氟苯乙胺合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=4:1(v/v)洗脱得到3′-N-[N-(2-氯-6-氟苯乙基)氨基硫代甲酰]十字孢碱(176)6.0mg,产率35.0%。[α]D 20+96°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.29(d,J=8.0Hz,1H,ArH),8.60(s,1H,NH),8.06(d,J=7.7Hz,1H,ArH),8.01(d,J=8.5Hz,1H,ArH),7.75(t,J=5.4Hz,1H,NH),7.69(d,J=8.3Hz,1H,ArH),7.48(m,1H,ArH),7.36(t,J=7.5Hz,1H,ArH),7.27-7.32(m,3H,ArH),7.15-7.19(m,1H,ArH),7.06(dd,J=8.3,6.6Hz,1H,H-1'),5.98(d,J=12.1Hz,1H,H-3'),5.00(s,2H,H-7),4.39(brs,1H,H-4'),3.89-3.96(m,1H,H-3”a),3.70-3.76(m,1H,H-3”b),3.03-3.16(m,2H,H-4”),2.77(s,3H,3'-NCH3),2.68(s,3H,4'-OCH3),2.67-2.71(m,1H,H-2'a),2.40(s,3H,6'-CH3),2.25(ddd,J=12.9,12.9,6.4Hz,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ181.8,172.0,161.4(d,1JC-F=246.5Hz),138.9,136.3,134.7(d,3JC-F=6.6Hz),132.7,129.3,128.9(d,3JC-F=10.4Hz),125.7,125.4×2,125.3(d,2JC-F=25.6Hz),125.1×2,123.8,122.7,121.5,120.4,119.5,119.4,115.2,114.3(d,2JC-F=22.3Hz),114.2,113.7,109.2,95.0,83.3,82.4,60.2,53.9,45.5,43.9,32.6,29.4,27.7,26.2;ESI-MS m/z 682.4/684.4[M+H]+.
化合物177的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(18.0mg,0.021mmol)与DMF、Et3N和2-间三氟甲基苯乙胺合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=4:1(v/v)洗脱得到3′-N-[N-(2-间三氟甲基苯乙基)氨基硫代甲酰]十字孢碱(177)6.0mg,产率35.0%。[α]D 20+132°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.29(d,J=8.0Hz,1H,ArH),8.61(s,1H,NH),8.05(d,J=7.8Hz,1H,ArH),7.99(d,J=8.5Hz,1H,ArH),,7.71(t,J=5.6Hz,1H,NH),7.70(d,J=8.0Hz,1H,ArH),7.52-7.58(m,4H,ArH),7.48(t,J=6.3Hz,2H,ArH),7.36(t,J=7.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),7.06(t,J=7.3Hz,1H,H-1'),5.95(d,J=12.4Hz,1H,H-3'),5.00(s,2H,H-7),4.44(brs,1H,H-4'),3.86-3.92(m,1H,H-3”a),3.74-3.80(m,1H,H-3”b),3.98-3.10(m,2H,H-4”),2.76(s,3H,3'-NCH3),2.70(s,3H,4'-OCH3),2.65-2.70(m,1H,H-2'a),2.38(s,3H,6'-CH3),2.24(ddd,J=12.9,12.9,6.8Hz,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ181.5,172.0,141.0,139.0,136.3,133.1,132.8,129.4,129.3(q,2JC-F=27.2Hz),128.9,125.7,125.4,125.2(q,3JC-F=5.0Hz),125.1,125.0,123.8,123.4(q,1JC-F=274.7Hz)122.9(q,3JC-F=5.7Hz),122.7,121.5,120.4,119.5,119.4,115.3,114.2,113.8,109.2,95.0,83.1,82.4,60.2,53.9,46.4,45.4,34.3,32.6,29.5,27.6;ESI-MS m/z 698.3[M+H]+.
化合物178的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(18.0mg,0.021mmol)与DMF、Et3N和苄胺合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=1:1(v/v)洗脱得到3′-N-(N-苯甲基氨基硫代甲酰)十字孢碱(178)9.0mg,产率73.2%。[α]D 20+54°(c0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.28(d,J=8.0Hz,1H,ArH),8.58(s,1H,NH),8.20(brs,1H,NH),8.05(d,J=7.9Hz,1H,ArH),8.00(d,J=8.3Hz,1H,ArH),7.70(d,J=8.0Hz,1H,ArH),7.48(t,J=7.8Hz,2H,ArH),7.36(t,J=7.8Hz,1H,ArH),7.28-7.33(m,5H,ArH),7.21-7.25(m,1H,ArH),7.06(t,J=7.4Hz,H-1'),5.98(d,J=13.5Hz,1H,H-3'),5.00(s,2H,H-7),4.89(d,J=5.6Hz,2H,H-3”),4.49(s,1H,H-4'),2.89(s,3H,3'-NCH3),2.72(s,3H,4'-OCH3),2.68-2.74(m,1H,H-2'a),2.38(s,3H,6'-CH3),2.24-2.30(m,1H,H-2'b);13CNMR(150MHz,DMSO-d6)δ182.2,172.0,139.8,139.0,136.4,132.8,129.2,128.2×2,127.2×2,126.6,125.7,125.4,125.1,125.1,123.8,122.7,121.5,120.4,119.6,119.4,115.3,114.2,113.8,109.2,95.1,83.2,82.5,60.4,54.3,48.4,45.6,32.9,29.5,27.8;ESI-MS m/z 616.3[M+H]+.
化合物179的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(18.0mg,0.021mmol)与DMF、Et3N和对甲氧基苄胺合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=1:1(v/v)洗脱得到3′-N-[N-(4-甲氧基苯甲基)氨基硫代甲酰]十字孢碱(179)9.0mg,产率69.8%。[α]D 20+40°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.28(d,J=7.9Hz,1H,ArH),8.58(s,1H,NH),8.13(t,J=5.4Hz,1H,NH),8.05(d,J=7.8Hz,1H,ArH),7.99(d,J=8.5Hz,1H,ArH),7.69(d,J=8.2Hz,1H,ArH),7.48(t,J=7.5Hz,2H,ArH),7.35(t,J=7.4Hz,1H,ArH),7.30(t,J=7.6Hz,1H,ArH),7.27(d,J=8.6Hz,2H,ArH),7.05(t,J=7.5Hz,1H,H-1'),6.88(d,J=8.6Hz,2H,ArH),5.00(s,2H,H-7),4.80(d,J=12.5Hz,1H,H-3'),4.47(brs,1H,H-4'),3.73(brs,2H,H-3”),2.86(s,3H,3'-NCH3),2.74(s,3H,4'-OCH3),2.67-2.73(m,1H,H-2'a),2.37(s,3H,6'-CH3),2.23-2.29(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ182.0,172.0,158.2,139.0,136.4,132.8,131.7,129.3,128.6×2,125.7,125.5,125.2,125.1,123.9,122.7,121.5,120.4,119.6,119.4,115.3,114.2,113.8,113.6×2,109.2,95.1,83.2,82.5,60.4,55.2,54.3,47.9,45.6,32.8,29.5,27.8;ESI-MS m/z 646.3[M+H]+.
化合物180的制备
按照化合物171的制备方法,由化合物167的咪唑部分的碘甲烷盐(15.0mg,0.021mmol)与DMF、Et3N和4-(2-哌嗪乙基)吗啉合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=1:3(v/v)洗脱得到3′-N-[4-(2-(4-吗啉)乙基)哌嗪甲酰]十字孢碱(180)8.0mg,产率55.1%。[α]D 20+72°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.29(d,J=7.9Hz,1H,ArH),8.58(s,1H,NH),8.06(d,J=7.5Hz,1H,ArH),8.00(d,J=8.5Hz,1H,ArH),7.59(d,J=8.3Hz,1H,ArH),7.50(t,J=7.6Hz,1H,ArH),7.47(t,J=7.7Hz,1H,ArH),7.36(t,J=7.5Hz,1H,ArH),7.30(t,J=7.7Hz,1H,ArH),6.97(dd,J=8.8,5.2Hz,1H,H-1'),5.00(s,2H,H-7),4.44(ddd,J=13.0,4.9,2.2Hz,1H,H-3'),4.36(brs,1H,H-4'),3.79(brs,4H,-N(CH2-CH2 )2O),3.33(brs,8H,-N(CH2 -CH2 )2N),3.14(brs,8H,piperazine-CH2 -CH2 -morpholine,-N(CH2 -CH2)2O),2.78-2.83(m,1H,H-2'a),2.71(s,3H,3'-NCH3),2.55(s,3H,4'-OCH3),2.39(s,3H,6'-CH3),2.28-2.34(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ172.4,163.6,139.0,136.7,133.0,130.0,126.1,126.0,125.8,125.6,124.2,123.1,122.0,120.9,119.9,119.8,115.4,114.5,113.7,109.4,95.1,84.4,82.7,64.2×2,60.3,52.3×2,51.8×2,51.3,51.1,50.6,45.9,44.4×2,33.4,29.4,27.8;ESI-MS m/z 708.4[M+H]+.
化合物181的制备
按照化合物171的制备方法,由化合物167的咪唑部分的碘甲烷盐(15.0mg,0.021mmol)与DMF、Et3N和2,6-二氟苯甲胺合成。快速柱色谱分离、二氯甲烷:乙酸乙酯=1:2(v/v)洗脱得到3′-N-[N-(2,6-二氟苯甲基)氨基甲酰]十字孢碱(181)9.0mg,产率67.6%。[α]D 20+48°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.28(d,J=7.9Hz,1H,ArH),8.60(s,1H,NH),8.04(d,J=7.7Hz,1H,ArH),7.95(d,J=8.6Hz,1H,ArH),7.65(d,J=8.2Hz,1H,ArH),7.45-7.48(m,2H,ArH),7.33-7.38(m,2H,ArH),7.29(t,J=7.5Hz,1H,ArH),7.06(t,J=7.8Hz,2H,ArH),6.98(t,J=7.6Hz,1H,H-1'),6.93(t,J=4.6Hz,1H,NH),4.99(s,2H,H-7),4.82(d,J=12.7Hz,1H,H-3'),4.34-4.44(m,2H,H-3”),4.21(brs,1H,H-4'),2.73(s,3H,3'-NCH3),2.61(s,3H,4'-OCH3),2.55-2.60(m,1H,H-2'a),2.31(s,3H,6'-CH3),2.16(ddd,J=13.0,13.0,6.7Hz,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ172.1,161.3×2(dd,1JC-F=247.5Hz,3JC-F=9.2Hz),157.7,139.1,136.4,132.8,129.5(t,3JC-F=11.0Hz),129.3,125.8,125.5,125.2,125.1,123.8,122.7,121.5,120.4,119.6,119.4,115.6(t,2JC-F=18.1Hz),115.3,114.2,113.8,111.5×2(d,2JC-F=19.9Hz),109.1,95.0,84.0,82.5,60.3,48.9,45.6,32.7,29.9,29.5,27.5;ESI-MS m/z 636.5[M+H]+.
化合物182的制备
按照化合物169的制备方法,由化合物168的咪唑部分的碘甲烷盐(50.0mg,0.070mmol)与DMF、Et3N和2S-羟基-1-丙胺合成。半制备HPLC分离、MeOH:H2O=4:1(v/v)洗脱得到3′-N-[N-(2S-羟基丙基)氨基硫代甲酰]十字孢碱(182)12.0mg,产率28.5%。[α]D 20+31°(c 0.07,CHCl3);1H NMR(600MHz,DMSO-d6)δ9.30(d,J=8.0Hz,1H,ArH),8.59(s,1H,NH),8.05(d,J=7.8Hz,1H,ArH),8.00(d,J=8.5Hz,1H,ArH),7.69(d,J=8.2Hz,1H,ArH),7.48(t,J=7.6Hz,1H,ArH),7.47(d,J=7.6Hz,1H,ArH),7.35(t,J=7.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),7.06(t,J=7.6Hz,1H,H-1'),5.99(d,J=12.8Hz,1H,H-3'),5.01(s,2H,H-7),4.75(s,1H,OH),4.45(s,1H,H-4'),3.97(brs,1H,H-4”),3.61-3.66(m,1H,H-3”a),3.40-3.46(m,1H,H-3”b),2.85(s,3H,3'-NCH3),2.71(s,3H,4'-OCH3),2.66-2.71(m,1H,H-2'a),2.39(s,3H,6'-CH3),2.24-2.30(m,1H,H-2'b),1.07(d,J=6.2Hz,3H,H-5”);13CNMR(150MHz,DMSO-d6)δ181.7,172.0,138.9,136.3,132.7,129.3,125.7,125.4,125.1×2,123.8,122.7,121.5,120.4,119.5,119.4,115.3,114.2,113.7,109.1,95.0,83.3,82.4,64.6,60.2,54.0,53.1,45.5,32.5,29.4,27.7,21.2;ESI-MS m/z 584.7[M+H]+.
化合物183的制备
将Fradcarbazole C(16.0mg,0.032mmol)用5mL甲醇溶解,加入100.0μL三乙胺和100mg盐酸羟胺,室温过夜反应。反应液用乙酸乙酯稀释,1N盐酸洗后无水硫酸钠干燥浓缩。半制备HPLC分离、MeOH:H2O=9:1(v/v)洗脱得到3′-N-(N-羟基氨基亚氨基甲基)十字孢碱(183)12.0mg,产率71.8%。[α]D 20+15°(c 0.07,MeOH);1H NMR(600MHz,DMSO-d6)δ10.32(s,1H,NH),9.27(d,J=7.9Hz,1H,ArH),8.63(s,1H,NH),8.30(s,1H,NH),8.05(d,J=7.8Hz,1H,ArH),7.94(d,J=8.5Hz,1H,ArH),7.70(d,J=8.2Hz,1H,ArH),7.50(t,J=7.3Hz,2H,ArH),7.36(t,J=7.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),6.98(t,J=7.6Hz,1H,H-1'),5.00(s,2H,H-7),4.47(brs,1H,H-3'),4.18(brs,1H,H-4'),3.16(s,1H,OH),2.93(s,3H,3'-NCH3),2.73(s,3H,4'-OCH3),2.70-2.75(m,1H,H-2'a),2.35(s,3H,6'-CH3),2.27-2.32(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ172.0,158.6,139.4,136.4,132.9,128.9,125.8,125.5,125.1,124.9,123.9,122.7,121.5,120.5,119.7,119.5,115.4,114.4,114.3,109.1,95.0,82.5,82.0,60.6,52.0,45.6,31.6,29.4,27.2;ESI-MS m/z 525.2[M+H]+.
化合物184的制备
将十字孢碱(46.6mg,0.1mmol)用5mL二氯甲烷溶解,依次加入加入催化量的DMAP、二环己基碳二亚胺(24.7mg,0.12mmol)和苯丁酸氮芥(36.4mg,0.12mmol)室温下反应2小时,倒入冰水中,二氯甲烷萃取,无水硫酸钠干燥后浓缩,硅胶柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得到3′-N-[4-[4-(N,N-二(2-氯乙基)氨基)苯基]丁酰]十字孢碱(184)60.1mg,产率80%。[α]D 20+99°(c 0.07,CHCl3);1H NMR(CD3OD)δ9.27(d,J=8.0Hz,1H,ArH),7.85(d,J=7.7Hz,1H,ArH),7.73(d,J=8.5Hz,1H,ArH),7.40-7.44(m,2H,ArH),7.27-7.30(m,2H,ArH),7.22(d,J=8.1Hz,1H,ArH),7.03(d,J=8.5Hz,2H,ArH),6.65(dd,J=9.0,4.9Hz,1H,H-1'),6.59(d,J=8.5Hz,2H,ArH),5.10-5.14(m,1H,H-3'),4.86-4.95(m,2H,H-7),3.95(brs,1H,H-4'),3.65(t,J=6.7Hz,4H,-N(CH2 -CH2Cl)2),3.57(t,J=6.7Hz,4H,-N(CH2-CH2 Cl)2),2.76(s,3H,3'-NCH3),2.56(t,J=7.9Hz,2H,H-2”),2.43(s,3H,4'-OCH3),2.43-2.49(m,1H,H-2'a),2.39(s,3H,6'-CH3),2.30-2.33(m,1H,H-2'b),1.90(t,J=7.8Hz,2H,H-4”),1.53-1.56(m,2H,H-3”);13C NMR(CD3OD)δ175.0,174.5,144.9,139.1,137.0,133.1,131.0,130.8,130.0×2,126.7,126.6,125.9,125.5,124.9,123.8,121.8,121.0,120.3,119.2,116.4,114.9,112.9,112.6×2,108.4,95.1,85.1,82.9,60.7,53.9×2,48.9,46.5,40.9×2,34.1,31.7,29.3,28.4,26.0,25.3;ESI-MS m/z752.2/754.3/756.2[M+H]+.
化合物185的制备
i)N-苄氧基甲基-2-(3-吲哚)-3-溴马来酰亚胺(185a)的制备
将2,3-二溴马来酰亚胺(2.55g,10.0mmol)置于100mL三口瓶中,用30mL干燥的DMF溶解,氮气保护,降至0℃,分两次加入氢化钠(480mg,12mmol,60%质量分数分散在石蜡中)。1h后加入10mL干燥的DMF溶解的氯甲基苄甲醚(2.08mL,15mmol),升至室温反应2h。后降至0℃,加入20mL饱和氯化铵溶液终止反应,乙酸乙酯萃取,无水硫酸钠干燥有机相后浓缩,加压柱色谱分离石油醚:乙酸乙酯=50:1(v/v)洗脱得化合物N-苄氧基甲基-2,3-二溴马来酰亚胺3.55g,产率94%。ESIMS m/z 395.4,397.3,399.5[M+Na]+。将镁丝(432mg,18.0mmol)置于100mL干燥的三口瓶中,加入4mL干燥的四氢呋喃,氩气置换,后无水导入溴乙烷(1.35mL,18.0mmol),室温反应15min后升至40℃反应30min。用导管引入吲哚(2.11g,18.0mmol),反应1h。后加入N-苄氧基甲基-2,3-二溴马来酰亚胺(3.29g,8.9mmol),室温反应4h。加入20mL饱和氯化铵溶液终止反应,乙酸乙酯萃取,无水硫酸钠干燥有机相后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得到化合物185a 3.46g,产率94%。1H-NMR(600MHz,DMSO-d6)δ8.93(brs,1H,NH),8.03(t,J=7.8Hz,1H,ArH),7.99(t,J=7.8Hz,1H,ArH),7.44(d,J=7.8Hz,1H,ArH),7.37(s,1H,ArH),7.27-7.36(m,5H,ArH),7.25(d,J=7.8Hz,1H,ArH),5.17(s,2H,PhCH2OCH2 N),4.67(s,2H,PhCH2 OCH2N);13C NMR(150MHz,DMSO-d6)δ168.8,166.2,137.8,137.8,136.6,131.5,128.2×2,127.6,127.5×2,124.5,122.6,122.4,120.6,114.1,112.4,103.7,70.5,67.5;ESIMS m/z 433.0,435.0[M+Na]+.
ii)N-苄氧基甲基-2-(1-叔丁氧羰基-3-吲哚)-3-溴马来酰亚胺(185b)的制备
将化合物185a(3.15g,8.39mmol)用50mL四氢呋喃溶解,加入3.81mL叔丁氧基甲酸酐(即二叔丁基二碳酸酯Boc2O,16.78mmol)和100mg对二甲氨基吡啶,室温反应4h。直接浓缩后柱色谱分离,石油醚:乙酸乙酯=20:1(v/v)洗脱得到化合物185b 3.90g,产率91%。1H-NMR(600MHz,CDCl3)δ8.23(s,1H,ArH),8.20(d,J=8.2Hz,1H,ArH),7.80(dd,J=7.3,8.2Hz,1H,ArH),7.41(t,J=7.3,1H,ArH),7.26(d,J=7.3Hz,1H,ArH),7.30-7.36(m,5H,ArH),5.17(s,2H,PhCH2OCH2 N),4.68(s,2H,PhCH2 OCH2N),1.71(s,9H,-C(CH3)3);13C NMR(150MHz,CDCl3)δ168.2,165.6,148.9,137.4,136.7,135.4,130.0,128.5×2,127.9,127.6×2,126.8,125.4,123.3,122.5,120.9,115.4,108.4,85.2,71.9,67.8,28.1×3;ESIMS m/z 511.2,513.0[M+H]+.
iii)N-苄氧基甲基-2-(1-叔丁氧羰基-3-吲哚)-3-(3-吲哚)马来酰亚胺(185c)的制备
将镁丝(366.7mg,15.32mmol)置于100mL干燥的三口瓶中,加入4mL干燥的四氢呋喃,氩气置换,后无水导入溴乙烷(1.15mL,15.32mmol),室温反应15min后升至40℃反应30min。用导管引入吲哚(1.8g,15.32mmol),反应1h。后加入化合物185b(3.9g,7.66mmol),室温反应4h。加入20mL饱和氯化铵溶液终止反应,乙酸乙酯萃取,无水硫酸钠干燥有机相后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得到化合物185c(3.98g,产率95%)。1H-NMR(600MHz,CDCl3)δ8.60(brs,1H,NH),8.19(d,J=7.8Hz,1H,ArH),8.09(s,1H,ArH),8.02(s,1H,ArH);7.81(t,J=7.8Hz,1H,ArH);7.29~7.41(m,5H,ArH);7.16(dt,J=8.0,3.0Hz,1H,ArH),7.10(t,J=8.2Hz,1H,ArH),7.05(d,J=7.8,1H,ArH),6.77~6.83(m,3H,ArH),5.23(s,2H,PhCH2OCH2 N),4.72(s,2H,PhCH2 OCH2N),1.68(s,9H,-C(CH3)3);13C NMR(150MHz,CDCl3)δ171.4,171.2,149.2,137.6,136.0,135.0,131.6,129.9,128.6,128.4×2,128.0,127.8,127.7×2,125.3,124.6,124.4,122.7,122.6,121.7,121.6,120.7,115.0,111.6,110.7,106.4,84.5,71.7,67.3,28.1×3;ESIMS m/z 546.2[M–H]–.
iv))6-O-三异丙基硅基-D-葡萄烯糖(185d)的制备
将0.206mL高氯酸缓慢加到40mL醋酸酐中,40℃下搅拌30min,后将温度降至30℃,将10g D-葡萄糖缓慢加入并搅拌30min。将反应液降温至10℃,3.1g赤磷,5.8mL液溴和3.6mL水依次缓慢加入,后升温至30℃继续搅拌2h。反应液用50mL冰水淬灭,乙酸乙酯萃取,酯层用无水硫酸钠干燥后浓缩。浓缩物用50mL乙酸乙酯溶解,降温至0℃,将16.1g锌粉,212mg CuSO4·5H2O和1.06g醋酸钠用130mL 60%的醋酸水溶液混匀,加入到反应液中。0℃反应1h后升至室温反应1h,将反应液过滤,后用乙酸乙酯萃取,酯层用无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得到10.4g 3,4,6-三(O-乙酰基)-D-葡萄烯糖(两步产率68%),ESIMS m/z 273.2[M+H]+。将5.2g 3,4,6-三(O-乙酰基)-D-葡萄烯糖(19.1mmol)用100mL甲醇溶解,加入300mg甲醇钠,室温反应1h,反应液用阳离子树脂调节pH为7,过滤后浓缩,加压柱色谱(乙酸乙酯洗脱)得到2.5g D-葡萄烯糖,产率90%;ESIMS m/z 147.1[M+H]+。将5.6g D-葡萄烯糖(38.4mmol)用100mL吡啶溶解,降温至0℃,加入11.34mL三异丙基氯硅烷(54.22mmol)和15.6g咪唑(230.4mmol),室温反应2h,用50mL冰水淬灭,乙酸乙酯萃取,酯层用无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=5:1(v/v)洗脱得到5.2g化合物185d,产率46%。1H-NMR(600MHz,CDCl3)δ6.30(d,J=6.0,1H,H-1),4.72-4.74(m,1H,H-2),4.27-4.29(m,1H,H-4),4.09(dd,J=12.0,4.8Hz,1H,H-6a),3.98(dd,J=12.0,4.8Hz,1H,H-6b),3.85(dd,J=6.0,3.6Hz,1H,H-3),3.81-3.84(1H,m,H-5),3.35(brs,1H,OH),2.35(brs,1H,OH),1.12-1.16(m,3H,-Si(CH(CH3)2)3),1.08(d,J=6.0Hz,18H,-Si(CH(CH 3)2)3).ESIMS m/z 303.2[M+H]+.
v)化合物185e的制备
将1.43g化合物185d(4.7mmol)置于100mL三口瓶中,用氩气保护,加入20mL干燥的二氯甲烷溶解,降至-5℃,分两次加入氢化钠(751mg,31.3mmol,60%分散在石蜡中),升至0℃反应20min,缓慢升至室温反应1.5h。重新降至-5℃,将三氯乙腈(5.59mL,56.4mmol)溶于10mL干燥的二氯甲烷中,用导管将其引入到反应液中,升至室温过夜反应。将反应液降至-78℃,滴加三氟化硼***(17.3mL,141mmol),在此温度下反应6h,后加入20mL饱和的碳酸氢钠溶液,缓慢升至室温,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=20:1(v/v)洗脱得1.1g化合物185e,产率52%。[α]D 20+121°(c 2.02,CH2Cl2);1H-NMR(600MHz,CDCl3)δ7.02(brs,1H,NH),6.45(d,J=4.8,1H,H-1),4.92-4.94(m,1H,H-2),4.46-4.48(m,1H,H-4),4.16-4.18(m,1H,H-3),4.06(dd,J=12.0,5.4Hz,1H,H-6a),3.96(dd,J=12.0,5.4Hz,1H,H-6b),3.84-3.86(m,1H,H-5),3.17(brs,1H,OH),1.12-1.16(m,3H,-Si(CH(CH3)2)3),1.08(d,J=6.0Hz,18H,-Si(CH(CH 3)2)3);13C-NMR(150MHz,CDCl3):δ162.3,145.8,97.3,92.6,74.5,67.1,63.4,45.8,17.8×3,11.7×6;ESIMS m/z 444.0[M–H]–.
vi)化合物185f的制备
将1.1g化合物185e(2.4mmol)溶于30mL二氯甲烷中,降至0℃,加入氢化钠(244mg,10.2mmol,60%分散在石蜡中),缓慢升至室温反应3h,后降至0℃,加入水终止反应,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得595mg化合物185f,产率75%。[α]D 20+108°(c 3.00,CH2Cl2);1H-NMR(500MHz,CDCl3)δ6.58(d,J=7.2,1H,H-1),5.96(brs,1H,NH),4.85-4.87(m,2H,H-2and H-4),4.34(1H,dd,J=7.2,4.2Hz,H-3),4.06(dd,J=10.2,3.6Hz,1H,H-6a),3.96(dd,J=10.2,3.6Hz,1H,H-6b),3.80-3.82(m,1H,H-5),1.12-1.16(m,3H,-Si(CH(CH3)2)3),1.08(d,J=6.0Hz,18H,-Si(CH(CH 3)2)3);13C-NMR(125MHz,CDCl3):δ158.7,147.2,98.5,74.0,71.0,61.7,46.1,17.9×3,11.9×6;ESIMS m/z 326.0[M–H]-.
vii)化合物185g的制备
将595mg化合物185f(1.8mmol)转入到两口瓶中,加入20mL二氯甲烷溶解,降至-5℃,加入氢化钠(218mg,9.1mmol,60%分散在石蜡中),升至室温反应两小时,后加入硫酸二甲酯(0.87mL,9.1mmol),室温反应16h,加入冰水终止反应,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=6:1(v/v)洗脱得600mg化合物185g,产率97%。[α]D 20+75°(c 1.00,CH2Cl2);1H-NMR(500MHz,CDCl3)δ6.68(d,J=7.2Hz,1H,H-1),4.93(1H,dd,J=7.2,4.8Hz,H-2),4.74-4.76(m,1H,H-4),4.09(dd,J=13.2,3.6Hz,1H,H-6a),4.07-4.10(m,1H,H-3),3.98(dd,J=13.2,3.6Hz,1H,H-6b),3.61-3.63(m,1H,H-5),2.84(s,3H,N-CH3),1.12-1.16(m,3H,-Si(CH(CH3)2)3),1.08(d,J=6.0Hz,18H,-Si(CH(CH 3)2)3);13C-NMR(125MHz,CDCl3)δ157.3,148.5,96.0,74.4,67.7,61.6,51.0,28.8,17.9×3,11.9×6.ESIMS m/z 342.2[M+H]+.
viii)混合物185h的制备
将415mg化合物185g(1.22mmol)溶于20mL四氢呋喃中,降至0℃,加入20mL水溶解的醋酸汞(781mg,2.44mmol),溶液变为黄色,升至室温反应2h。降至0℃,加入60mL水,后缓慢加入硼氢化钠(371mg,9.76mmol),有黑色生成,10min后通入二氧化碳至溶液呈中性。抽滤后用乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得到一对未分离的C-1位差向异构体185h(337mg,产率77%)。1H-NMR(600MHz,CDCl3)δ5.31(m,1H,H-1),5.15(dd,J=6.0,4.8Hz,1H,H-1),4.63(t,J=8.4Hz,1H,H-4),4.57(t,J=8.4Hz,1H,H-4),4.03-3.97(m,2H,H-6a),3.91-3.94(m,2H,H-6b),3.86-3.90(m,2H,H-3),3.78-3.81(m,1H,H-5),3.58-3.61(m,1H,H-5),2.86(s,3H,N-CH3),2.83(s,3H,N-CH3),2.22-2.26(m,1H,H-2a),2.04-2.08(m,1H,H-2a),1.96-2.01(m,1H,H-2b),1.81(ddd,J=13.2,8.4,6.0Hz,1H,H-2b),1.07-1.12(m,6H,-Si(CH(CH3)2)3),1.05(d,J=7.2Hz,36H,-Si(CH(CH 3)2)3);13C-NMR(150MHz,CDCl3)δ158.4,158.0,91.7,90.6,74.7,69.0,68.8,68.1,63.2,63.1,53.9,52.9,31.2,29.9,29.2,28.9,18.0×3,17.8×3,12.4×6,12.0×6;ESIMS m/z 360.2[M+H]+.
ix)化合物185i和185j的制备
将712.5mg化合物185c(1.253mmol)置于250mL三口反应瓶中,加入20mL干燥的四氢呋喃溶解,氩气保护,降温至-78℃,加入10mL干燥的四氢呋喃溶解的三苯基膦(655mg,2.515mmol),后将0.5mL DIAD(2.515mmol)溶于10mL四氢呋喃中,滴加入反应液,在-78℃反应1h后加入10mL四氢呋喃溶解的化合物185h(300mg,0.835mmol),在-78℃反应2h后升至室温过夜反应。加入饱和的氯化铵溶液终止反应,乙酸乙酯萃取后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得到202mg化合物185i(收率27%)210mg化合物185j(收率28%)。
185i:[α]D 20+14.1°(c 0.59,CH2Cl2);1H NMR(600MHz,CDCl3)δ8.15(d,J=7.8Hz,1H,ArH),8.11(s,1H,ArH),7.77(s,1H,ArH),7.40-7.10(m,9H,ArH),6.86(t,J=7.8Hz,1H,ArH),6.80-6.77(m,2H,ArH),5.72(dd,J=10.8,1.8Hz,1H,H-1'),5.23(s,2H,PhCH2OCH2 N),4.59(dd,J=9.0,7.2Hz,1H,H-4'),4.72(s,2H,PhCH2 OCH2N),4.06-4.08(m,1H,H-3'),4.01(dd,J=12.0,1.8Hz,1H,H-6'a),3.95(dd,J=12.0,2.4Hz,1H,H-6'b),3.82-3.84(m,1H,H-5'),2.87(s,3H,N-CH3),2.39-2.41(m,1H,H-2'a),2.27-2.30(m,1H,H-2'b),1.69(s,9H,-C(CH 3)3),1.04-1.10(m,3H,-Si(CH(CH3)2)3),1.02(d,J=6.0Hz,-Si(CH(CH 3)2)3);13C NMR(150MHz,CDCl3)δ171.1,171.0,158.5,149.1,137.6,135.7,135.1,130.5,129.0,128.6,128.3×2,127.7,127.6×2,126.5,125.7,124.6,123.2,122.4×2,121.7,121.5,121.0,115.1,110.6,110.4,107.0,84.6,79.4,78.3,71.7,67.3,67.0,62.9,55.7,29.5,29.3,28.1×3,17.8×3,11.8×6;ESIMS m/z 889.6[M+H]+,911.6[M+Na]+;HR-ESIMS m/z889.4195[M+H]+(calcd for C50H61N4O9Si,889.4208).
185j:[α]D 20-9.1°(c 0.10,CH2Cl2);1H NMR(600MHz,CDCl3)δ8.15(d,J=7.2Hz,1H,ArH),8.13(s,1H,ArH),7.63(s,1H,ArH),7.39-7.42(m,3H,ArH),7.28-7.32(m,3H,ArH),7.22-7.25(m,1H,ArH),7.15-7.19(m,2H,ArH),6.95(t,J=7.2Hz,1H,ArH),6.78(t,J=7.8Hz,1H,ArH),6.70(d,J=7.8Hz,1H,ArH),6.10(dd,J=10.7,4.5Hz,1H,H-1'),5.23(s,2H,PhCH2OCH2 N),4.75(t,J=7.8Hz,1H,H-4'),4.72(s,2H,PhCH2 OCH2N),3.99-4.03(m,1H,H-3'),3.85-3.93(m,2H,H-6'),3.80-3.83(m,1H,H-5'),2.87(s,3H,N-CH3),2.40-2.44(m,1H,H-2'a),2.06-2.11(m,1H,H-2'b),1.69(s,9H,-C(CH 3)3),1.06-1.12(m,3H,-Si(CH(CH3)2)3),1.02(d,J=6.0Hz,18H,-Si(CH(CH 3)2)3);13C NMR(150MHz,CDCl3)δ171.2,171.0,157.3,149.2,137.8,136.0,135.4,130.8,129.4,128.5×3,127.9,127.8×2,127.3,126.7,126.0,124.7,123.3,122.5×2,122.0,121.7,115.4,110.6,110.4,107.1,84.8,78.6,72.5,71.8,68.8,67.4,63.6,53.7,29.8,29.2,28.2×3,18.0×3,11.9×6.ESIMS m/z 889.5[M+H]+.
x)化合物185l和188b的制备
将化合物185i(311mg,0.350mmol)用40mL甲苯溶解,加入3.0g硅胶,加热回流5h。降至室温后用硅胶过滤,乙酸乙酯洗脱得到化合物185k(262mg,产率95%);HR-ESIMS m/z787.3499[M–H]-(calcd for C45H51N4O7Si,787.3572)。以化合物185j(210mg)为原料,以同样的方法制得188a(180mg,产率96%),HR-ESIMS m/z 787.3496[M–H]-(calcd forC45H51N4O7Si,787.3572)。将262mg化合物185k(0.333mmol)用30mL四氢呋喃溶解,降至0℃,加入四丁基氟化铵(1.0mL,1.0mmol,1.0M四氢呋喃溶液)脱保护基,室温反应1h,乙酸乙酯稀释后水洗,乙酸乙酯层用无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得到179mg化合物185l,产率85%。以188a(179mg)为原料,以同样的方法制得188b(136mg,产率95%)。
185l:[α]D 20-2.5°(c 0.01,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.40(s,1H,NH),7.67(d,J=3.0Hz,1H,ArH),7.59(s,1H,ArH),7.36-7.37(m,2H,ArH),7.27-7.32(m,4H,ArH),7.22-7.24(m,1H,ArH),7.17-7.17(m,2H,ArH),7.04(t,J=7.8Hz,1H,ArH),6.87-6.90(m,2H,ArH),6.76(t,J=7.8Hz,1H,ArH),5.67(dd,J=10.2,1.2Hz,1H,H-1'),5.14(s,2H,PhCH2OCH2 N),4.67(s,2H,PhCH2 OCH2N),4.30(t,J=7.8Hz,1H,H-4'),3.86-3.87(m,1H,H-3'),3.81(dd,J=12.0,2.4Hz,1H,H-6'a),3.72-3.75(m,1H,H-5'),3.64(dd,J=12.0,2.4Hz,1H,H-6'b),2.77(s,3H,N-CH3),2.24-2.29(m,1H,H-2'a),2.12-2.16(m,1H,H-2'b);13C NMR(150MHz,CDCl3):δ171.7,171.6,158.6,137.7,136.3,136.0,129.7,129.2,128.5×2,128.0,127.8×2,127.7,126.6,126.5,124.9,123.3,122.9,122.6,122.2,121.5,120.4,111.8,110.0,107.6,106.5,78.9,77.6,71.7,67.2,62.1,60.5,55.7,29.6,28.8;HR-ESIMS m/z 631.2216[M-H]-(calcd for C36H31N4O7,631.2193).
188b:[α]D 20-10°(c 0.10,CH2Cl2);1H NMR(600MHz,CDCl3)δ8.90(s,1H,NH),7.80(m,1H,ArH),7.46-7.21(m,10H,ArH),7.80(d,J=3.0Hz,1H,ArH),7.45-7.46(m,2H,ArH),7.38-7.40(m,3H,ArH),7.34-7.36(m,1H,ArH),7.29-7.32(m,2H,ArH),7.20-7.25(m,2H,ArH),7.07-7.10(m,1H,ArH),7.01(t,J=7.2Hz,1H,ArH),6.72-6.74(m,2H,ArH),6.05(dd,J=10.4,5.4Hz,1H,H-1'),5.14(s,2H,PhCH2OCH2 N),4.71(s,2H,PhCH2 OCH2N),4.60(t,J=8.4Hz,1H,H-4'),3.92-3.97(brs,1H,H-3'),3.69(dd,J=12.0,2.4Hz,1H,H-6'a),3.60(dd,J=12.0,4.2Hz,1H,H-6'b),3.52-3.54(m,1H,H-5'),2.80(s,3H,N-CH3),2.39-2.43(m,1H,H-2'a),2.25-2.30(m,1H,H-2'b);13C NMR(150MHz,CDCl3):δ171.7×2,157.3,137.8,136.4,135.8,129.7,129.1,128.5×2,127.9,127.8×2,127.7,127.0,126.8,124.3,123.3,123.0,122.7,122.3,121.6,120.4,112.0,110.1,107.6,106.5,78.4,76.5,71.8,68.9,62.1,59.2,53.2,29.8,28.9;HR-ESIMS m/z 631.2218[M-H]-(C36H31N4O7,计算值631.2193).
xi)化合物185m和188c的制备
将30mg化合物185l(0.047mmol)溶于1750mL丙酮中,加入3mg碘催化,250w高压汞灯照射反应12h,溶液由红色变为绿色荧光,浓缩后加入饱和硫代硫酸钠溶液,乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得产物185m(17mg,产率57%)。以化合物188b(136mg)为原料,以相同的方法制得化合物188c(70mg,产率51%)。
185m:[α]D 20+71.7°(c 0.05,CH2Cl2);1H NMR(600MHz,CDCl3)δ11.0(s,1H,NH),9.23(d,J=7.8Hz,1H,ArH),8.77(d,J=7.8Hz,1H,ArH),7.54-7.57(m,1H,ArH),7.46-7.48(m,2H,ArH),7.39-7.41(m,2H,ArH),7.35-7.38(m,2H,ArH),7.28-7.31(m,3H,ArH),6.97(d,J=8.4Hz,1H,ArH),6.16(dd,J=9.0,1.8Hz,1H,H-1'),5.18(t,J=7.8Hz,1H,H-4'),5.13-5.17(m,2H,PhCH2OCH2 N),4.80(s,2H,PhCH2 OCH2N),4.40(d,J=12.0Hz,1H,H-3'),4.10-4.13(m,2H,H-6'),3.35-3.38(m,1H,H-5'),2.91(s,3H,N-CH3),2.29-2.34(m,1H,H-2'a),1.98-2.01(m,1H,H-2'b);13C NMR(150MHz,CDCl3):δ169.5,169.0,158.7,140.7,139.7,137.5,129.7,128.6×2,128.3,128.1×3,128.0,127.6,127.2,126.2,125.0,122.7,121.7,120.7,120.5,119.5,118.4,118.1,111.2,108.6,79.3,78.3,71.9,66.8,66.2,60.9,56.2,29.7,29.0;ESIMS m/z 629.3[M-H]-.
188c:[α]D 20-25.7°(c 0.16,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ11.4(s,1H,NH),9.22(d,J=7.8Hz,1H,ArH),9.10(d,J=7.8Hz,1H,ArH),7.95(d,J=8.4Hz,1H,ArH),7.79(d,J=8.4Hz,1H,ArH),7.73-7.74(m,1H,ArH),7.67-7.68(m,1H,ArH),7.61-7.64(m,2H,ArH),7.46(t,J=7.8Hz,1H,ArH),7.40(t,J=7.2Hz,1H,ArH),7.32(d,J=7.2Hz,1H,ArH),7.31(t,J=7.8Hz,1H,ArH),7.24(t,J=7.2Hz,1H,ArH),6.94(dd,J=11.4,4.8Hz,1H,H-1'),5.45(t,J=5.4Hz,1H,H-4'),5.20(s,2H,PhCH2OCH2 N),4.83-4.85(m,1H,H-3'),4.77-4.79(m,1H,OH),4.69(s,2H,PhCH2 OCH2N),4.44-4.48(m,1H,H-5'),3.90-3.96(m,2H,H-6'),2.67(s,3H,N-CH3),2.48-2.52(m,1H,H-2'a),2.43-2.46(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ169.6,169.5,157.2,141.2,140.0,138.4,132.3,132.1,129.3×2,129.1,128.8×2,128.1×3,128.0,125.3,124.9,122.6,122.0,121.5 121.3,120.3,118.8,118.2,112.8,79.5,75.1,70.1,67.3,65.6,61.4,53.4,29.6,29.0;ESIMS m/z 629.2[M-H]-.
xii)化合物185o和188e的制备
将307mg三苯基膦(1.171mmol)和159mg咪唑(2.342mmol)用20mL二氯甲烷溶解,降至0℃,加入287mg碘(2.342mmol),搅拌1h。将化合物185m(123mg,0.195mmol)用20mL二氯甲烷溶解,缓慢加入到反应液中,升至室温反应6h。后降至0℃,加入水猝灭,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得到化合物185n(80mg,产率56%);HR-ESIMS m/z 741.1221[M+H]+(C36H30N4O6I,计算值741.1210)。以化合物188c(70mg)为原料,以相同的方法制得化合物188d(54mg,产率65%),HR-ESIMS m/z741.1225[M+H]+(calcd for C36H30N4O6I,741.1210)。将30mg化合物185n(0.041mmol)用10mL四氢呋喃溶解,降至0℃,加入0.4mL DBU(2.67mmol),0℃反应1h,升至40℃反应1h。反应液用乙酸乙酯稀释,水洗后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得到化合物185o(22.3mg,产率89%)。以化合物188d(54mg)为原料,以相同的方法制得化合物188e(40mg,产率90%)。
185o:[α]D 20+141.3°(c 0.34,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ12.1(s,1H,NH),9.22(d,J=7.8Hz,1H,ArH),9.08(d,J=8.4Hz,1H,ArH),7.86(d,J=8.4Hz,1H,ArH),7.71(d,J=8.4Hz,1H,ArH),7.60-7.63(m,2H,ArH),7.47(d,J=7.2Hz,1H,ArH),7.41-7.36(m,3H,ArH),7.31(t,J=7.2Hz,2H,ArH),7.24(t,J=7.2Hz,1H,ArH),7.20(dd,J=12.0,2.4Hz,1H,H-1'),5.40(d,J=9.6,1H,H-4'),5.15-5.19(m,2H,PhCH2OCH2 N),5.07-5.09(m,2H,H-6'),4.67(s,2H,PhCH2 OCH2N),4.33-4.36(m,1H,H-3'),2.69(s,3H,N-CH3),2.44-2.51(m,1H,H-2'a),2.10-2.14(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ169.6,169.5,157.0,152.9,142.2,140.6,138.4,129.8,129.4,128.8×2,128.2,128.1×3,128.0,125.6,124.9,123.7,122.5,121.8,121.4,120.5,119.2,118.8,118.4,113.7,112.9,101.2,81.0,71.4,70.9,67.3,53.1,28.8,28.0;ESIMS m/z 611.3[M-H]-.
188e:[α]D 20-21.4°(c 0.7,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.62(s,1H,NH),9.27(d,J=8.4Hz,1H,ArH),9.10(d,J=8.4Hz,1H,ArH),7.62-7.66(m,1H,ArH),7.51-7.56(m,2H,ArH),7.38-7.46(m,4H,ArH),7.29-7.36(m,3H,ArH),7.22(t,J=7.2Hz,1H,ArH),6.27(dd,J=11.4,2.4Hz,1H,H-1′),5.43(d,J=2.0Hz,1H,H-6′a),5.30(d,J=2.0Hz,1H,H-6′b),5.16-5.22(m,2H,PhCH2OCH2 N),5.07(d,1H,J=7.2Hz,H-4′),4.73(s,2H,PhCH2 OCH2N),4.14-4.18(m,1H,H-3′),2.76(s,3H,N-CH3),2.47-2.53(m,1H,H-2′a),2.38-2.42(m,1H,H-2′b);13C NMR(150MHz,CDCl3)δ169.4,169.2,156.7,151.3,140.8,139.7,137.7,129.3,128.5×2,128.2,128.0×3,127.8,127.5,126.3,125.3,122.6,122.2,121.9,121.6,121.0,119.5,119.3,118.8,111.9,108.7,100.1,81.9,71.6,70.1,66.9,54.4,32.8,29.2;ESIMS m/z 611.4[M-H]-.
xiii)化合物185p和188f的制备
将30mg化合物185o(0.05mmol)用四氢呋喃/甲醇10mL/1mL溶解,降温至0℃,加入22mg叔丁醇钾(0.2mmol),溶液由黄色变为红色,缓慢升至室温搅拌2h,加入38mg碘(0.15mmol),溶液颜色加深,过夜反应。降温至0℃,倒入到饱和硫代硫酸钠溶液中,乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得产物185p(20mg,产率54%)。以化合物188e(40mg)为原料,以相同的方法制得化合物188f(20mg,产率42%)。
185p:[α]D 20+45.9°(c 0.05,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ9.28(d,J=7.8Hz,1H,ArH),9.03(d,J=7.8Hz,1H,ArH),8.11(d,J=9.0Hz,1H,ArH),8.07(d,J=8.4Hz,1H,ArH),7.70-7.66(m,2H,ArH),7.56(t,J=7.2Hz,1H,ArH),7.47(t,J=7.2Hz,1H,ArH),7.36-7.40(m,2H,ArH),7.31(t,J=7.2Hz,2H,ArH),7.24(t,J=7.2Hz,1H,ArH),7.08(dd,J=9.6,7.2Hz,1H,H-1'),5.91(d,J=12.0Hz,1H,H-4'),5.26(s,2H,PhCH2OCH 2),4.70(s,2H,PhCH 2OCH2),4.63(1H,d,J=12.6Hz,H-6′a),4.48-4.51(m,1H,H-3′),3.79(1H,d,J=12.6Hz,H-6′b),2.99(s,3H,N-CH3),2.58-2.63(m,1H,H-2′a),2.36-2.42(m,1H,H-2′b);13CNMR(150MHz,DMSO-d6)δ169.2,169.0,156.7,138.1,135.5,132.4,131.0,128.9,128.5×2,127.9,127.8×2,127.5,126.2,126.0,125.4,122.7,121.9,121.4,119.7,119.1,118.5,117.9,116.7,112.5,108.0,93.5,77.7,71.8,70.5,67.0,53.2,29.8,27.8,9.3;ESIMS m/z761.1[M+Na]+.
188f:[α]D 20-73.4°(c 0.21,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.41(d,J=8.4Hz,1H,ArH),9.23(d,J=8.4Hz,1H,ArH),8.05(d,J=8.4Hz,1H,ArH),7.61(t,J=7.2Hz,2H,ArH),7.49(t,J=7.2Hz,1H,ArH),7.43-7.46(m,4H,ArH),7.30(t,J=7.2Hz,2H,ArH),7.22(t,J=7.2Hz,1H,ArH),6.63(dd,J=10.8,6.0Hz,1H,H-1′),5.34(s,2H,PhCH2OCH 2),5.31(d,J=9.0Hz,1H,H-4′),4.73(s,2H,PhCH 2OCH2),4.52(d,J=11.4Hz,1H,H-6′a),4.27-4.32(m,1H,H-3′),3.96(1H,d,J=11.4Hz,H-6′b),2.84-2.89(m,1H,H-2′a),2.80(s,3H,N-CH3),2.41-2.48(m,1H,H-2′b);13C NMR(150MHz,CDCl3)δ169.4,169.2,155.4,141.7,137.8×2,131.8,128.7,128.4×2,128.0,127.9×2,127.8,127.6,126.6,126.1,125.1,122.8,122.3,121.7,121.3,119.7×2,115.0,114.1,107.1,92.2,79.4,73.5,71.6,67.0,53.4,29.5,29.0,14.2;ESIMS m/z 761.1[M+Na]+.
vix)化合物185q和188g的制备
将25mg化合物185p(0.034mmol)溶于20mL苯中,氩气保护,加入AIBN(3mg)和四丁基氢化锡(0.1mL),加热回流1h。降至室温后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得产物185q(17mg,产率80%)。以化合物188f(20mg)为原料,以相同的方法制得化合物188g(15mg,产率96%)。
185q:[α]D 20+87.3°(c 0.29,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.35(d,J=7.2Hz,1H,ArH),9.10(d,J=7.8Hz,1H,ArH),7.64-7.57(m,3H,ArH),7.47-7.42(m,4H,ArH),7.35(t,J=7.2Hz,1H,ArH),7.31(t,J=7.8Hz,2H,ArH),7.23(t,J=7.2Hz,1H,ArH),6.49(dd,J=9.6,7.2Hz,1H,H-1'),5.63(d,J=9.6Hz,1H,H-4'),5.25-5.33(m,2H,PhCH2OCH 2),4.76(s,2H,PhCH 2OCH2),4.32-4.35(m,1H,H-3'),3.10(s,3H,N-CH3),2.69-2.73(m,1H,H-2'a),2.36-2.40(m,1H,H-2'b),1.94(s,3H,6'-CH3);13C NMR(150MHz,CDCl3)δ169.4,169.2,157.3,140.1,138.1,137.7,130.1,128.9,128.5×2,128.0×2,127.8,127.6,127.5,126.7,126.3,124.7,122.2,122.1,121.5,121.0,119.6,118.5,117.5,112.3,107.8,94.0,77.2,71.7,71.4,66.9,52.7,29.6,26.2,24.6;ESIMS m/z 613.5[M+H]+.
188g:[α]D 20-24.0°(c 0.19,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.40(d,J=7.8Hz,1H,ArH),9.24(d,J=7.8Hz,1H,ArH),8.09(d,J=7.8Hz,1H,ArH),7.72-7.74(m,1H,ArH),7.59-7.62(m,2H,ArH),7.53-7.55(m,1H,ArH),7.43-7.47(m,3H,ArH),7.31(t,J=7.8Hz,2H,ArH),7.25(t,J=7.2Hz,1H,ArH),6.59(dd,J=9.6,6.6Hz,1H,H-1'),5.37(s,2H,PhCH2OCH 2),5.11(d,J=8.4Hz,1H,H-4'),4.77(s,2H,PhCH 2OCH2),4.26-4.31(m,1H,H-3'),2.79-2.85(m,1H,H-2'a),2.77(s,3H,N-CH3),2.42-2.48(m,1H,H-2'b),2.08(s,3H,6'-CH3);13C NMR(150MHz,CDCl3)δ169.5,169.3,155.8,142.1,137.7×2,131.0,128.9,128.7,128.4×2,127.9×2,127.8,127.7,127.6,126.6,125.8,124.6,122.3,122.2,121.6,119.7,119.5,116.4,114.1,107.5,93.3,79.0,71.9,71.6,67.0,53.0,30.0,29.7,29.5;ESIMS m/z 635.2[M+Na]+.
xx)化合物185和188的制备
将化合物185q(10mg,0.016mmol)溶于20mL乙酸乙酯:甲醇=1:1(v/v)中,氩气置换后加入5mg 20%的氢氧化钯碳,后氢气置换,过夜反应。用硅胶过滤后浓缩,半制备HPLC分离、MeOH:H2O=9:1(v/v)洗脱得4′-O-去甲基-(4′-O,3′-N)羰基-7-氧亚基-3′,4′-双表十字孢碱(185)7mg,产率89%。以化合物188g(15mg)为原料,以相同的方法制得对映[4′-O-去甲基-(4′-O,3′-N)羰基-7-氧亚基]十字孢碱(188)11.3mg,产率96%。
185:[α]D 20+33.5°(c 0.04,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ11.2(s,1H,NH),9.22(d,J=7.8Hz,1H,ArH),9.00(d,J=7.8Hz,1H,ArH),8.08(d,J=7.8Hz,1H,ArH),7.95(d,J=7.8Hz,1H,ArH),7.65(t,J=7.8Hz,1H,ArH),7.63(t,J=7.2Hz,1H,ArH),7.49(t,J=7.8Hz,1H,ArH),7.42(t,J=7.8Hz,1H,ArH),6.99(dd,J=10.2,6.6Hz,1H,H-1'),5.76(d,J=9.6Hz,1H,H-4'),4.43-4.46(m,1H,H-3'),2.99(s,3H,N-CH3),2.80(ddd,J=9.6,6.0,2.4Hz,1H,H-2'a),2.18(ddd,J=9.6,6.0,2.4Hz,1H,H-2'b),1.80(s,3H,6'-CH3);13CNMR(150MHz,DMSO-d6)δ171.2×2,157.4,140.4,135.9,130.2,129.1,127.8,127.7,125.8,125.3,124.3,122.0×2,121.5,121.4,120.7,117.6,116.5,114.4,114.3,94.1,77.9,71.4,52.4,29.5,24.9×2;ESIMS m/z 491.3[M-H]-.
188:[a]D 20-50.2°(c 0.08,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ9.23(d,J=9.0Hz,1H,ArH),9.04(d,J=10.2Hz,1H,ArH),8.09(d,J=9.6Hz,1H,ArH),7.88(d,J=10.8Hz,1H,ArH),7.64(t,J=9.0Hz,1H,ArH),7.57-7.60(m,1H,ArH),7.41-7.44(m,2H,ArH),7.01(dd,J=12.0,7.2Hz,1H,H-1'),5.33(d,J=10.8Hz,1H,H-4'),4.32-4.36(m,1H,H-3'),2.93-2.99(m,1H,H-2'a),2.57(s,3H,N-CH3),2.04-2.11(m,1H,H-2'b),2.05(s,3H,6'-CH3);13CNMR(150MHz,DMSO-d6)δ170.8,170.5,155.7,141.4,137.7,130.0,128.2,127.3,126.7,125.0,124.7,123.5,121.3,121.1×2,121.0,119.9,117.5,116.7,115.8,109.4,92.6,79.0,75.3,52.0,29.8,28.3×2;ESIMS m/z 515.2[M+Na]+.
化合物186,187,189,190的制备
将化合物185(15mg,0.030mmol)溶于10mL甲醇中,降至0℃加入硼氢化钠(7.6mg,0.2mmol),升至室温反应两小时,溶液由黄色变为无色,用乙酸乙酯稀释后加入饱和氯化铵溶液,乙酸乙酯萃取,无水硫酸钠干燥后蒸干。将粗产物溶于5mL冰醋酸中,加入锌粉(20mg,0.32mmol),升至40℃反应1.5小时,降至室温后,用乙酸乙酯稀释,饱和碳酸氢钠溶液洗涤后,无水硫酸钠干燥后浓缩。半制备HPLC色谱分离、乙腈:水=2:3(v/v)洗脱得5.2mg 4′-O-去甲基-(4′-O,3′-N)羰基-3′,4′-双表十字孢碱(186)(产率36%)和5.2mg 5-脱氧-7-氧亚基-4′-O-去甲基-(4′-O,3′-N)羰基-3′,4′-双表十字孢碱(187)(产率36%)。以化合物188(11mg)为原料,以相同的方法制得4.0mg对映[4′-O-去甲基-(4′-O,3′-N)羰基]十字孢碱(189)(产率为38%)和4.0mg对映[5-脱氧-7-氧亚基-4′-O-去甲基-(4′-O,3′-N)羰基]十字孢碱(190)(产率为38%)。
186:[α]D 20+101.0°(c 0.10,MeOH);1H NMR(600MHz,CDCl3)δ9.32(d,J=8.0Hz,1H,ArH),7.97(d,J=7.7Hz,1H,ArH),7.68(d,J=8.3Hz,1H,ArH),7.56(m,1H,ArH),7.52-7.58(m,2H,ArH),7.43-7.48(m,2H,ArH),7.38(t,J=7.5Hz,1H,ArH),6.57(s,1H,NH),6.49(dd,J=9.6,6.8Hz,1H,H-1'),5.59(d,J=9.4Hz,1H,H-4'),5.04(d,J=16.7Hz,1H,H-7a),4.92(d,J=16.7Hz,1H,H-7b),4.28-4.30(m,1H,H-3'),3.06(s,3H,N-CH3),2.49-2.55(m,1H,H-2'a),2.22-2.28(m,1H,H-2'b),1.94(s,3H,6'-CH3);13C NMR(125MHz,CDCl3)δ172.7,157.4,139.2,137.2,133.2,129.3,127.2,126.3,125.8×2,125.5,123.5,122.1,121.8,120.8,120.3,117.8,115.8,113.0,107.5,93.9,77.2,71.7,52.9,46.1,29.6,26.2,24.7;HR-ESIMS m/z 479.1738[M+H]+(calcd for C28H23N4O4 479.1719).
187:[α]D 20+102.6°(c 0.05,MeOH);1H NMR(600MHz,CDCl3)δ9.60(d,J=7.7Hz,1H,ArH),7.98(d,J=7.7Hz,1H,ArH),7.61(d,J=8.4Hz,1H,ArH),7.56(t,J=7.7Hz,1H,ArH),7.54(t,J=8.4Hz,1H,ArH),7.51(d,J=8.4Hz,1H,ArH),7.44(t,J=7.3Hz,1H,ArH),7.40(t,J=7.4Hz,1H,ArH),6.50(dd,J=9.4,6.9Hz,1H,H-1'),6.40(s,1H,NH),5.67(d,J=9.5Hz,1H,H-4'),4.99-5.07(m,2H,H-5),4.30(d,J=9.5Hz,1H,H-3'),3.08(s,3H,N-CH3),2.59-2.64(m,1H,H-2'a),2.37-2.43(m,1H,H-2'b),1.90(s,3H,6'-CH3);13C NMR(150MHz,CDCl3)δ173.1,157.5,139.3,137.3,134.7,128.2,127.7,126.8,126.2×2,125.9,123.2,122.0,121.5,121.1,119.6,118.9,115.3,111.9,108.4,94.0,77.2,71.8,52.9,45.7,29.6,26.4,24.4;HR-ESIMS m/z 479.1706[M+H]+(calcd for C28H23N4O4,479.1719).
189:[α]D 20-72.8°(c 0.10,MeOH);1H NMR(600MHz,DMSO-d6)δ9.23(d,J=7.6Hz,1H,ArH),8.67(s,1H,NH),8.06(d,J=8.5Hz,1H,ArH),8.03(d,J=7.4Hz,1H,ArH),7.79(d,J=8.4Hz,1H,ArH),7.50-7.53(m,2H,ArH),7.37-7.40(m,1H,ArH),7.29-7.32(m,1H,ArH),6.96(dd,J=9.9,6.3Hz,1H,H-1'),5.31(d,J=8.8Hz,1H,H-4'),4.95-5.03(m,2H,H-7),4.34(ddd,J=12.1,8.8,5.1Hz,1H,H-3'),2.90-2.94(m,1H,H-2'a),2.58(s,3H,N-CH3),2.03(s,3H,6'-CH3),1.97-2.02(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ171.7,155.7,140.4,136.5,133.0,128.7,125.8,125.6,125.1,125.0,124.7,122.4,121.3,121.1,120.4,119.7,116.7,115.9,115.5,108.8,92.6,79.2,75.5,52.1,45.4,29.6,28.8,28.3;HR-ESIMS m/z 479.1708[M+H]+(calcd for C28H23N4O4 479.1719).
190:[α]D 20-75.6°(c 0.10,MeOH);1H NMR(600MHz,DMSO-d6)δ9.51(d,J=7.9Hz,1H,ArH),8.62(s,1H,NH),8.10(d,J=7.8Hz,1H,ArH),8.01(d,J=8.5Hz,1H,ArH),7.84(d,J=8.2Hz,1H,ArH),7.56(t,J=7.7Hz,1H,ArH),7.45-7.48(m,1H,ArH),7.38(t,J=7.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),6.96(dd,J=9.8,6.3Hz,1H,H-1'),5.29(d,J=8.7Hz,1H,H-4'),4.96-5.01(m,2H,H-5),4.34(ddd,J=12.1,8.8,5.1Hz,1H,H-3'),2.91-2.96(m,1H,H-2'a),2.59(3H,s,N-CH3),2.05-2.10(m,1H,H-2'b),1.99(s,3H,6'-CH3);13CNMR(150MHz,DMSO-d6)δ171.9,155.8,140.4,136.6,134.4,127.0,126.6,125.70,125.5,125.1×2,122.1,121.9,120.6,120.1,119.4,117.6,116.1,114.3,109.3,92.6,79.2,75.5,52.2,44.9,29.5,28.8,28.3;HR-ESIMS m/z 479.1722[M+H]+(calcd for C28H23N4O4479.1719).
化合物191的制备
i)6-O-三异丙基硅基-L-葡萄烯糖(191a)的制备
将0.041mL高氯酸缓慢加到40mL醋酸酐中,40℃下搅拌30min,后将温度降至30℃,加入2g L-葡萄糖缓慢并继续搅拌30min。将反应液降温至10℃,依次缓慢加入0.62g赤磷、1.16mL液溴和0.72mL水,升温至30℃继续搅拌2h。用10mL冰水淬灭反应,乙酸乙酯萃取,无水硫酸钠干燥后浓缩。浓缩物用50mL乙酸乙酯溶解,降温至0℃,将3.22g锌粉、42.4mgCuSO4·5H2O和0.21g醋酸钠用30mL 60%体积分数的醋酸水溶液混匀后加入到反应液中,0℃反应1h后升至室温反应1h。将反应液过滤,滤液用乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=3:1(v/v)洗脱得到2.5g 3,4,6-三(O-乙酰基)-L-葡萄烯糖,两步产率82%;ESIMS m/z 273.3[M+H]+。将3,4,6-三(O-乙酰基)-L-葡萄烯糖(2.5g,9.19mmol)用100mL甲醇溶解,加入60mg甲醇钠,室温反应1h,反应液用阳离子树脂调节pH为7,过滤后浓缩,加压柱色谱分离(乙酸乙酯洗脱)得到1.3g L-葡萄烯糖,产率97%;ESIMS m/z 147.2[M+H]+。化合物L-葡萄烯糖(1.3g,8.9mmol)用30mL吡啶溶解,降温至0℃,加入三异丙基氯硅烷(3.78ml,17.8mmol)和咪唑(3.61g,53.4mmol),室温反应2h,用50mL冰水淬灭,乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=5:1(v/v)洗脱得到化合物191a 1.41g,产率52%。1H-NMR(500MHz,CDCl3)δ6.27(d,J=6.0Hz,1H,H-1),4.67-4.70(m,1H,H-2),4.26(brs,1H,H-4),4.04(dd,J=12.0,4.8Hz,1H,H-6a),3.98(dd,J=12.0,4.8Hz,1H,H-6b),3.79-3.82(m,2H,H-3and H-5),1.09-1.16(m,3H,((CH3)2CH)3Si-),1.06(d,J=6.0Hz,18H,((CH 3)2CH)3Si-).13C-NMR(125MHz,CDCl3)δ143.9,102.5,76.5,72.5,69.4,64.4,17.8×3,11.8×6;ESIMS m/z 325.1[M+Na]+.
ii)化合物191b的制备
将1.41g化合物191a(4.7mmol)置于100mL三口瓶中,氩气保护下加入20mL干燥的二氯甲烷搅溶,降至-5℃,分两次加入氢化钠(740mg,18.5mmol,60%质量分数分散在石蜡中),升至0℃反应20min,缓慢升至室温继续反应1.5h。重新降至-5℃,将三氯乙腈(5.51mL,55.6mmol)溶于10mL干燥的二氯甲烷中,用导管将其引入到反应液中,升至室温过夜反应。将反应液降至-78℃,滴加三氟化硼***(17.0mL,139mmol),在此温度下反应6h,后加入20mL饱和碳酸氢钠溶液淬灭反应,缓慢升至室温,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=20:1(v/v)洗脱得到0.88g化合物191b,产率42%。[α]D 20-71.6°(c 0.90,CH2Cl2);1H-NMR(500MHz,CDCl3)δ7.02(brs,1H,NH),6.45(d,J=6.0,1H,H-1),4.93(dd,J=6.0,5.0Hz,1H,H-2),4.46-4.49(m,1H,H-4),4.16-4.18(m,1H,H-3),4.06(dd,J=12.0,5.4Hz,1H,H-6a),3.96(dd,J=12.0,5.4Hz,1H,H-6b),3.82-3.86(m,1H,H-5),1.10-1.17(m,3H,((CH3)2CH)3Si-),1.07(d,J=6.0Hz,18H,((CH 3)2CH)3Si-);13C-NMR(125MHz,CDCl3)δ162.4,145.8,97.3,92.6,74.6,67.3,63.5,45.9,17.9×3,11.8×6;ESIMS m/z446.1/448.2/490.1[M+H]+.
iii)化合物191c的制备
将化合物191b(0.88g,1.98mmol)溶于30mL二氯甲烷中,降至0℃,加入氢化钠(194mg,4.95mmol,60%分散在石蜡中),缓慢升至室温反应3h,后降至0℃,加入水终止反应,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得537mg产物191c,产率84%。[α]D 20-86.0°(c 0.50,CH2Cl2);1H-NMR(500MHz,CDCl3)δ6.59(d,J=6.0,1H,H-1),5.87(brs,1H,NH),4.85-4.88(m,2H,H-2/H-4),4.34(dd,J=7.5,4.0Hz,1H,H-3),4.04(dd,J=11.0,3.0Hz,1H,H-6a),3.99(dd,J=11.0,3.60Hz,1H,H-6b),3.82-3.84(m,1H,H-5),1.09-1.16(m,3H,((CH3)2CH)3Si-),1.06(d,J=6.0Hz,18H,((CH 3)2CH)3Si-);13C-NMR(125MHz,CDCl3)δ158.5,147.2,98.5,74.1,71.1,61.8,46.1,17.9×3,11.9×6;ESIMS m/z 326.1[M–H]–.
iv)化合物191d的制备
将化合物191c(537mg,1.64mmol)转入到两口瓶中,加入20mL二氯甲烷溶解,降至-5℃,加入氢化钠(197mg,4.92mmol,60%分散在石蜡中),升至室温反应两小时,后加入硫酸二甲酯(0.79mL,8.21mmol),室温反应16h,加入冰水终止反应,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=6:1(v/v)洗脱得467mg化合物191d,产率83%。[α]D 20-77°(c 1.00,CH2Cl2);1H-NMR(500MHz,CDCl3)δ6.67(d,J=6.0Hz,1H,H-1),4.93(dd,J=6.0,4.0Hz,1H,H-2),4.74(1H,dd,J=9.0,7.5Hz,H-4),4.05-4.07(m,1H,H-3),4.04(dd,J=12.0,2.5Hz,1H,H-6a),4.00(dd,J=12.0,2.5Hz,1H,H-6b),3.60-3.63(1H,m,H-5),2.84(s,3H,N-CH3)1.08-1.15(m,3H,((CH3)2CH)3Si-),1.06(d,J=6.0Hz,18H,((CH 3)2CH)3Si-);13C-NMR(125MHz,CDCl3)δ157.3,148.4,96.0,74.5,67.8,61.7,51.0,28.6,17.9×3,11.9×6;ESIMS m/z 342.2[M+H]+.
v)化合物191e的制备
将化合物191d(467mg,1.37mmol)溶于20mL四氢呋喃中,降至0℃,加入20mL水溶解的醋酸汞(876mg,2.74mmol),溶液变为黄色,升至室温反应2h。降至0℃,加入60mL水,后缓慢加入硼氢化钠(416mg,11.0mmol),10min后通入二氧化碳至溶液呈中性。抽滤后用乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得到400mg一对1-位差向的异构体混合物191e,产率81%。1H-NMR(500MHz,CDCl3):δ5.30-5.33(m,1H,H-1),5.15-5.18(m,1H,H-1),4.64-4.68(m,1H,H-4),4.556-4.60(m,1H,H-4),4.04-3.86(m,6H,H-3,H-6a,H-6b,),3.77-3.82(m,1H,H-5),3.60-3.63(m,1H,H-5),2.87(s,3H,N-CH3),2.84(s,3H,N-CH3),2.22-2.27(m,1H,H-2a),2.05-2.09(m,1H,H-2a),1.97-2.02(m,1H,H-2b),1.79-1.85(m,1H,H-2b),1.08-1.15(m,6H,((CH3)2CH)3Si-),1.07(d,J=7.2Hz,36H,((CH 3)2CH)3Si-).13C-NMR(125MHz,CDCl3)δ158.2,157.8,91.6,90.6,74.5,69.0,68.7,68.1,63.2,63.0,53.7,52.8,31.1,29.9,29.1,28.8,17.9×6,12.0×12;ESIMS m/z360.2[M+H]+.
vi)化合物191f和191g的制备
将870mg化合物185c(1.59mmol)置于250mL三口反应瓶中,加入20mL干燥的四氢呋喃溶解,氩气保护,降温至-78℃,加入10mL干燥的四氢呋喃溶解的三苯基膦(833mg,3.18mmol),后将0.62mL DIAD(3.18mmol)溶于10mL四氢呋喃中,滴加入反应液,在–78℃反应1h后加入10mL四氢呋喃溶解的化合物191e(380mg,1.06mmol),在-78℃反应2h后升至室温过夜反应。加入饱和的氯化铵溶液终止反应,乙酸乙酯萃取后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=4:1(v/v)洗脱得到226mg化合物191f(收率24%)和230mg化合物191g(收率25%)。
191f:[α]D 20-14.2°(c 1.0,CH2Cl2);1H NMR(500MHz,CDCl3)δ8.15(d,J=7.8Hz,1H,ArH),8.11(s,1H,ArH),7.77(s,1H,ArH),7.36-7.40(m,3H,ArH),7.29-7.32(m,2H,ArH),7.23-7.25(m,1H,ArH),7.15-7.18(m,1H,ArH),7.10-7.12(m,2H,ArH),6.86(t,J=7.8Hz,1H,ArH),6.77-6.81(m,2H,ArH),5.71(dd,J=10.5,2.0Hz,1H,H-1'),5.20-5.25(m,2H,PhCH2OCH 2N),4.71(s,2H,PhCH 2OCH2N),4.69(dd,J=9.0,7.5Hz,1H,H-4'),4.05-4.09(1H,m,H-3'),4.00(dd,J=12.0,2.0Hz,1H,H-6'a),3.95(dd,J=12.0,2.0Hz,1H,H-6'b),3.82-3.85(1H,m,H-5'),2.88(s,3H,N-CH3),2.39-2.44(m,1H,H-2'a),2.27-2.30(m,1H,H-2'b),1.69(s,9H,(CH 3)3CO-),1.05-1.11(m,3H,((CH3)2CH)3Si-),1.02(d,J=6.0Hz,18H,((CH 3)2CH)3Si-).13C NMR(125MHz,CDCl3)δ171.1,171.0,158.5,149.2,137.7,135.8,135.2,130.6,129.0,128.6,128.4×2,127.7×2,127.6×2,126.6,125.8,124.6,123.2,122.5,122.2,121.8,121.6,115.1,110.6,110.5,107.1,84.6,79.5,78.4,71.7,67.4,67.1,63.0,55.8,29.6,29.4,28.2×3,17.9×3,11.9×6;ESIMS m/z 889.6[M+H]+
191g:[α]D 20+9.3°(c 0.40,CH2Cl2);1H NMR(500MHz,CDCl3)δ8.15(d,J=7.2Hz,1H,ArH),8.13(s,1H,ArH),7.64(s,1H,ArH),7.39-7.42(m,3H,ArH),7.28-7.33(m,3H,ArH),7.22-7.25(m,1H,ArH),7.15-7.20(m,2H,ArH),6.95(t,J=7.0Hz,1H,ArH),6.78(t,J=8.0Hz,1H,ArH),6.72(d,J=8.0Hz,1H,ArH),6.10(dd,J=10.0,6.0Hz,1H,H-1'),5.23(s,2H,PhCH2OCH 2N),4.76(t,J=7.8Hz,1H,H-4'),4.72(s,2H,PhCH 2OCH2N),3.99-4.04(m,1H,H-3'),3.85-3.94(m,2H,H-6'),3.80-3.83(m,1H,H-5'),2.87(s,3H,N-CH3),2.39-2.45(m,1H,H-2'a),2.05-2.13(m,1H,H-2'b),1.69(s,9H,(CH 3)3CO-),1.05-1.10(m,3H,((CH3)2CH)3Si-),1.02(d,J=6.0Hz,18H,((CH 3)2CH)3Si-).13C NMR(125MHz,CDCl3)δ171.0,170.9,157.2,149.2,137.8,136.0,135.4,130.8,129.3,128.4×3,127.7,127.6×2,127.3,126.7,126.0,124.6,123.2,122.4×2,121.9,121.6,115.3,110.5,110.3,107.1,84.7,78.6,72.4,71.8,68.8,67.4,63.5,53.6,29.7,29.2,28.2×3,17.9×3,11.9×6;ESIMS m/z 889.6[M+H]+.
vii)化合物191i和194b的制备
将化合物191f(226mg,0.254mmol)用40mL甲苯溶解,加入3.0g硅胶,加热回流5h。降至室温后用硅胶过滤,乙酸乙酯洗脱得到200mg化合物191h,产率100%;ESIMS m/z787.4[M+H]+。以化合物191g(226mg)为原料,以同样的方法制得化合物194a(201mg,产率100%);ESIMS m/z 787.5[M+H]+。将化合物191h(200mg,0.253mmol)用30mL四氢呋喃溶解,降至0℃,加入四丁基氟化铵(1.0mL,1.0mmol,1.0M的THF溶液),室温反应1h,乙酸乙酯稀释后水洗,乙酸乙酯层用无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得到158mg化合物191i,产率98%。以194a(201mg)为原料,以同样的方法制得152mg化合物194b,产率94%。
191i:[α]D 20+11.6°(c 0.55,CH2Cl2);1H NMR(500MHz,CDCl3)δ8.82(brs,1H,NH),7.69(d,J=3.0Hz,1H,ArH),7.62(s,1H,ArH),7.37-7.38(m,2H,ArH),7.28-7.33(m,4H,ArH),7.23-7.26(m,1H,ArH),7.15-7.19(m,1H,ArH),7.14(d,J=7.5Hz,1H,ArH),7.07-7.10(m,1H,ArH),6.-6.92(m,2H,ArH),6.77-6.80(m,1H,ArH),5.71(dd,J=10.5,2.0Hz,1H,H-1'),5.17(s,2H,PhCH2OCH2 N),4.68(s,2H,PhCH2 OCH2N),4.46(1H,dd,J=9.0,7.0Hz,H-4'),3.92-3.95(m,1H,H-3'),3.85-3.87(m,1H,H-6'a),3.76-3.79(m,1H,H-5'),3.66-3.69(m,1H,H-6'b),2.82(s,3H,N-CH3),2.30-2.36(m,1H,H-2'a),2.20-2.23(m,1H,H-2'b).13C NMR(125MHz,CDCl3)δ171.6,171.5,158.4,137.7,136.1,136.0,129.2,129.0,128.4×2,127.8,127.7×2,127.6,126.8,126.5,124.8,123.2,122.8,122.7,122.2,121.4,120.4,111.5,109.8,107.6,106.7,78.8,77.6,71.6,67.2,62.1,60.5,55.7,29.5,28.9;ESIMS m/z 631.3[M-H]-.
194b:[α]D 20+19.3°(c 0.25,CH2Cl2);1H NMR(500MHz,CDCl3)δ8.90(s,1H,NH),7.83(d,J=3.0Hz,1H,ArH),7.47(s,1H,ArH),7.45(d,J=7.5Hz,1H,ArH),7.38-7.41(m,3H,ArH),7.36(d,J=8.0Hz,1H,ArH),7.29-7.32(m,2H,ArH),7.20-7.25(m,2H,ArH),7.08-7.11(m,1H,ArH),7.00-7.03(t,J=7.2Hz,1H,ArH),6.73-6.76(m,2H,ArH),6.06(dd,J=10.0,5.0Hz,1H,H-1'),5.20-5.25(m,2H,PhCH2OCH2 N),4.72(s,2H,PhCH2 OCH2N),4.62(t,J=9.5Hz,1H,H-4'),3.94-3.99(m,1H,H-3'),3.71-3.73(m,1H,H-6'a),3.61-3.63(m,1H,H-6'b),3.54-3.57(m,1H,H-5'),2.81(s,3H,N-CH3),2.42-2.47(m,1H,H-2'a),2.01-2.08(m,1H,H-2'b);13C NMR(125MHz,CDCl3)δ171.5×2,157.0,137.8,136.2,135.8,129.3,129.0,128.4×2,127.7×3,127.4,127.2,126.8,124.3,123.2,122.8,122.7,122.2,121.5,120.4,111.6,109.8,107.7,106.7,78.3,71.7,71.3,68.6,67.3,62.1,53.2,29.7,28.9;ESIMS m/z:631.3[M-H]-.
viii)化合物191j和194c的制备
将10mg化合物191i(0.016mmol)溶于1000mL丙酮中,加入1mg碘催化,125w高压汞灯照射反应12h,溶液由红色变为绿色荧光,浓缩后加入饱和硫代硫酸钠溶液,乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得6.1mg产物191j,产率61%。以化合物194b(10mg)为原料,以相同的方法制得5.3mg化合物194c,产率53%。
191j:[α]D 20-51.7°(c 0.07,CH2Cl2);1H NMR(600MHz,CDCl3):δ11.0(s,1H,NH),9.06(d,J=7.8Hz,1H,ArH),8.64(d,J=7.8Hz,1H,ArH),7.47(t,J=7.7Hz,1H,ArH),7.39(d,J=7.5Hz,2H,ArH),7.38(d,J=7.7Hz,1H,ArH),7.31(t,J=7.8Hz,2H,ArH),7.23-7.26(m,3H,ArH),7.19(t,J=7.8Hz,1H,ArH),6.85(t,J=8.4Hz,1H,ArH),6.11(d,J=11.0Hz,1H,H-1'),5.15(t,J=7.8Hz,1H,H-4'),4.95(s,2H,PhCH2OCH2 N),4.70(s,2H,PhCH2 OCH2N),4.28(d,J=10.8Hz,1H,H-3'),4.04-4.08(m,3H,H-5',H-6'),2.88(s,3H,N-CH3),2.21-2.26(m,1H,H-2'a),1.97-2.00(m,1H,H-2'b);13C NMR(150MHz,CDCl3):δ169.2,168.9,158.8,140.7,139.6,137.5,129.6,128.5×2,128.1,127.9×3,127.4,126.9,125.9,124.7,122.4,121.5,121.4,120.4,120.2,119.2,118.1,117.8,111.3,108.6,79.1,78.3,71.6,66.5,66.3,60.5,56.1,29.6,28.9;ESIMS m/z 629.3[M-H]-.
194c:[α]D 20+32.6°(c 0.11,CH2Cl2);1H NMR(600MHz,CDCl3):δ9.78(s,1H,NH),8.91-8.94(m,2H,ArH),7.47(d,J=8.2Hz,1H,ArH),7.39-7.41(m,3H,ArH),7.34(t,J=7.5Hz,1H,ArH),7.31(t,J=7.8Hz,2H,ArH),7.27(d,J=8.2Hz,1H,ArH),7.23-7.26(m,1H,ArH),7.21(t,J=7.8Hz,1H,ArH),7.16(t,J=7.8Hz,1H,ArH),6.62-6.64(m,1H,H-1'),5.00-5.08(m,2H,PhCH2OCH2 N),4.70(s,2H,PhCH2 OCH2N),4.68(brs,1H,H-4'),4.49-4.50(m,1H,H-3'),4.31-4.33(m,1H,H-5'),4.04-4.09(m,2H,H-6'),2.59(s,3H,N-CH3),2.07-2.11(m,1H,H-2'a),1.97-2.03(m,1H,H-2'b);13C NMR(150MHz,CDCl3)δ169.2,169.0,157.0,140.6,139.7,137.6,129.1,128.6×2,128.1×2,128.0×2,127.7,127.1,125.8,124.9,122.2,121.6×2,121.0,123.1,118.7,118.3,118.1,111.9,109.0,77.6,75.3,70.9,66.8,64.6,60.5,54.6,29.8,28.8;ESIMS m/z 629.2[M-H]-.
ix)化合物191l和194e的制备
将242mg三苯基膦(0.92mmol)和咪唑(126mg,1.85mmol)用20mL二氯甲烷溶解,降至0℃,加入碘(234mg,1.85mmol),搅拌1h。将化合物191j(97mg,0.15mmol)用20mL二氯甲烷溶解,缓慢加入到反应液中,升至室温反应6h。后降至0℃,加入水猝灭,二氯甲烷萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=2:1(v/v)洗脱得到105mg化合物191k,产率77%;ESIMS m/z 741.3[M+H]+。以化合物194c(80mg)为原料,以相同的方法制得化合物194d(90mg,产率95%);ESIMS m/z 741.2[M+H]+。将化合物191k(105mg,0.14mmol)用10mL四氢呋喃溶解,降至0℃,加入DBU(0.4mL,2.67mmol),0℃反应1h,升至40℃反应1h。反应液用乙酸乙酯稀释,水洗后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:1(v/v)洗脱得到80mg化合物191l,产率92%。以化合物194d(90mg)为原料,以相同的方法制得化合物194e(67mg,产率90%)。
191l:[α]D 20-96.0°(c 0.55,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ12.2(s,1H,NH),9.22(d,J=7.8Hz,1H,ArH),9.08(d,J=8.4Hz,1H,ArH),7.88(d,J=8.4Hz,1H,ArH),7.75(d,J=8.4Hz,1H,ArH),7.64(m,2H,ArH),7.49(d,J=7.2Hz,1H,ArH),7.34-7.43(m,5H,ArH),7.29(m,1H,ArH),7.24(m,1H,H-1'),5.46(d,J=8.5Hz,1H,H-4'),5.13(s,2H,PhCH2OCH2 N),5.10(m,2H,H-6'),4.67(s,2H,PhCH2 OCH2N),4.40(m,1H,H-3'),2.77(3H,s,N-CH3),2.53(m,1H,H-2'a),2.18(m,1H,H-2'b);13C NMR(150MHz,DMSO-d6)δ169.4,169.3,157.1,152.9,142.1,140.5,138.3,129.6,129.2,128.8×2,128.2,128.1×3,127.9,125.5,124.8,123.6,122.4,121.7,121.4,120.3,119.2,118.6,118.4,113.6,112.8,101.2,81.0,71.4,70.9,67.0,53.0,28.8,28.1;ESIMS m/z 611.1[M-H]-.
194e:[α]D 20+12.4°(c 0.20,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.62(s,1H,NH),9.28(d,J=8.2Hz,1H,ArH),9.10(d,J=8.2Hz,1H,ArH),7.66(d,J=8.2Hz,1H,ArH),7.55-7.59(m,2H,ArH),7.42-7.47(m,4H,ArH),7.36-7.41(m,3H,ArH),7.30(t,J=7.8Hz,1H,ArH),6.30(dd,J=11.4,2.4Hz,1H,H-1′),5.52(d,J=2.0Hz,1H,H-6′a),5.36(d,J=2.0Hz,1H,H-6′b),5.11(d,1H,J=7.2Hz,H-4′),5.08-5.18(m,2H,PhCH2OCH2 ),4.67(s,2H,PhCH2 OCH2N),4.18-4.21(m,1H,H-3′),2.83(s,3H,N-CH3),2.51-2.57(m,1H,H-2′a),2.43-2.47(m,1H,H-2′b);13C NMR(150MHz,CDCl3)δ169.3,169.2,156.7,151.1,140.9,139.9,138.0,129.4,128.6×2,128.3,128.1×2,128.0,127.9,127.7,126.4,125.4,122.7,122.4,121.9,121.7,121.0,119.5,119.3,118.9,112.0,108.9,100.2,82.0,71.7,70.3,66.9,54.6,33.0,29.3;ESIMS m/z 611.4[M-H]-.
x)化合物191m和194f的制备
将化合物191l(80mg,0.13mmol)用四氢呋喃/甲醇10mL/1mL溶解,降温至0℃,加入叔丁醇钾(59mg,0.2mmol),溶液由黄色变为红色,缓慢升至室温搅拌2h,加入碘(133mg,0.15mmol),溶液颜色加深,过夜反应。降温至0℃,倒入到饱和硫代硫酸钠溶液中,乙酸乙酯萃取,无水硫酸钠干燥后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得60mg产物191m,产率62%。以化合物194e(40mg)为原料,以相同的方法制得化合物194f(25mg,产率52%)。
191m:[α]D 20-40.2°(c 0.06,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ9.22(d,J=7.8Hz,1H,ArH),9.00(d,J=7.8Hz,1H,ArH),8.10(d,J=8.4Hz,1H,ArH),8.05(d,J=8.4Hz,1H,ArH),7.63-7.67(m,2H,ArH),7.49-7.54(m,1H,ArH),7.43(t,J=7.2Hz,1H,ArH),7.37-7.38(m,2H,ArH),7.29-7.32(m,2H,ArH),7.23(t,J=7.2Hz,1H,ArH),7.09(dd,J=9.6,7.2Hz,1H,H-1'),5.92(d,J=12.0Hz,1H,H-4'),5.20(s,2H,PhCH2OCH 2),4.69(s,2H,PhCH 2OCH2),4.61(1H,d,J=12.6Hz,H-6′a),4.51-4.53(m,1H,H-3′),3.80(1H,d,J=12.6Hz,H-6′b),3.01(s,3H,N-CH3),2.86-2.90(m,1H,H-2′a),2.51-2.55(m,1H,H-2′b);13CNMR(150MHz,DMSO-d6)δ169.4,169.1,156.8,139.8,138.5,138.2,137.4,132.0,129.1,128.6×2,128.1,127.9×3,125.5,125.0,124.4,122.3,121.5,121.2,120.5,119.2,117.5,116.8,113.0,110.6,93.3,78.2,70.8,70.3,65.4,52.7,29.3,28.4,11.6;ESIMS m/z 761.1[M+Na]+.
194f:[α]D 20+51.3°(c 0.12,CH2Cl2);1H NMR(600MHz,DMSO-d6)δ9.25(d,J=8.2Hz,1H,ArH),9.03(d,J=8.2Hz,1H,ArH),8.17(d,J=8.2Hz,1H,ArH),7.90(d,J=8.2Hz,1H,ArH),7.66(m,1H,ArH),7.65-7.68(m,1H,ArH),7.60-7.63(m,1H,ArH),7.44-7.50(m,2H,ArH),7.36(d,J=8.2Hz,2H,ArH),7.30(t,J=7.2Hz,2H,ArH),7.23(t,J=7.2Hz,1H,ArH),7.04-7.07(m,1H,H-1′),5.49(d,J=9.0Hz,1H,H-4′),5.13-5.18(m,2H,PhCH2OCH 2),4.93(d,J=11.4Hz,1H,H-6′a),4.67(s,2H,PhCH 2OCH2),4.34-4.38(m,1H,H-3′),3.96(1H,d,J=11.4Hz,H-6′b),2.99-3.03(m,1H,H-2′a),2.65(s,3H,N-CH3),2.51-2.55(m,1H,H-2′b);13C NMR(150MHz,DMSO-d6)δ169.6,169.3,155.9,141.6,138.4,138.3,132.2×2,132.1,128.9,128.8×2,128.1×4,125.4,125.1,124.4,122.3,121.8,121.5,120.5,119.2,118.2,116.7,116.1,110.2,92.6,80.1,73.6,71.0,65.6,53.1,30.6,29.0,14.2;ESIMS m/z 761.1[M+Na]+.
xi)化合物191n和194g的制备
将化合物191m(60mg,0.08mmol)溶于20mL苯中,氩气保护,加入AIBN(10mg)和四丁基氢化锡(0.2mL),加热回流1h。降至室温后浓缩,加压柱色谱分离、石油醚:乙酸乙酯=1:2(v/v)洗脱得42mg产物191n,产率85%。以化合物194f(25mg)为原料,以相同的方法制得化合物194g(20mg,产率96%)。
191n:[α]D 20-63.5°(c 0.09,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.30(d,J=7.2Hz,1H,ArH),9.10(d,J=7.8Hz,1H,ArH),7.68-7.61(m,3H,ArH),7.51–7.43(m,4H,ArH),7.37(t,J=7.2Hz,3H,ArH),7.30(t,J=7.8Hz,1H,ArH),6.55(dd,J=9.6,7.2Hz,1H,H-1'),5.71(d,J=9.6Hz,1H,H-4'),5.20-5.31(m,2H,PhCH2OCH 2),4.81(s,2H,PhCH 2OCH2),4.44-4.46(m,1H,H-3'),3.18(s,3H,N-CH3),2.76-2.81(m,1H,H-2'a),2.46-2.51(m,1H,H-2'b),1.99(s,3H,6'-CH3);13C NMR(150MHz,CDCl3)δ169.3,169.1,157.3,140.1,138.1,137.7,130.1,128.9,128.5×2,127.9×2,127.8,127.6,127.4,126.5,126.2,124.6,122.1,122.0,121.4,120.9,119.4,118.4,117.4,112.3,107.8,94.0,77.2,71.7,71.4,66.8,52.7,29.6,26.2,24.6;ESIMS m/z 613.5[M+H]+.
194g:[α]D 20+33.1°(c 0.13,CH2Cl2);1H NMR(600MHz,CDCl3)δ9.38(d,J=8.0Hz,1H,ArH),9.21(d,J=7.9Hz,1H,ArH),8.08(d,J=8.6Hz,1H,ArH),7.57-5.60(m,2H,ArH),7.41-7.46(m,5H,ArH),7.31(t,J=7.6Hz,2H,ArH),7.23(t,J=7.4Hz,1H,ArH),6.58(dd,J=10.3,6.4Hz,1H,H-1'),5.30-5.35(m,2H,PhCH2OCH 2),5.09(d,J=8.9Hz,1H,H-4'),4.76(s,2H,PhCH 2OCH2),4.24-4.28(m,1H,H-3'),2.80-2.84(m,1H,H-2'a),2.76(s,3H,3'-NCH3),2.41-2.47(m,1H,H-2'b),2.07(s,3H,6'-CH3);13C NMR(150MHz,CDCl3)δ169.5,169.3,155.9,142.2,137.9,137.8,130.7,128.7,128.5×2,128.0×2,127.8,127.6,127.5,126.5,125.8,124.6,122.3,122.2,121.6,120.9,119.7,119.5,117.3,116.4,107.6,93.3,79.1,76.2,71.6,67.0,53.1,30.1,29.8,29.7;ESIMS m/z 613.6[M+H]+.
xii)化合物191和194的制备
将化合物191n(40mg,0.065mmol)溶于乙酸乙酯/甲醇(10mL/10mL)共20mL中,氩气置换后加入20mg 20%的氢氧化钯碳,后氢气置换,过夜反应。用硅胶过滤后浓缩,半制备HPLC分离、MeOH:H2O=7:3(v/v)洗脱得到28mg 4′-O-去甲基-(4′-O,3′-N)羰基-7-氧亚基-1′,5′-双表十字孢碱(191),产率89%。以化合物194g(20mg)为原料,以相同的方法制得15mg 4′-O-去甲基-(4′-O,3′-N)羰基-7-氧亚基十字孢碱(194),产率96%。
191:[α]D 20-69.5°(c 2.0,CH2Cl2);1H NMR(500MHz,DMSO-d6)δ11.2(s,1H,NH),9.21(d,J=7.8Hz,1H,ArH),8.98(d,J=7.8Hz,1H,ArH),8.07(d,J=7.8Hz,1H,ArH),7.94(d,J=8.4Hz,1H,ArH),7.61-7.67(m,2H,ArH),7.48(t,J=7.8Hz,1H,ArH),7.41(t,J=7.8Hz,1H,ArH),6.98-7.01(m,1H,H-1'),5.76(d,J=10.4Hz,1H,H-4'),4.44-4.46(m,1H,H-3'),2.99(s,3H,N-CH3),2.81(ddd,J=9.6,6.0,2.4Hz,1H,H-2'a),2.18(ddd,J=9.6,6.0,2.4Hz,1H,H-2'b),1.80(s,3H,6'-CH3);13C NMR(125MHz,DMSO-d6)δ171.5,171.2,157.4,140.5,138.6,130.2,129.1,127.8,127.7,125.8,125.3,124.3,122.1,122.0,121.5,121.4,120.7,117.6,116.5,114.3,110.7,94.1,77.9,71.4,52.4,29.5,25.4,24.9;ESIMS m/z 491.3[M-H]-.
194:[α]D 20+112.9°(c 0.1,CH2Cl2);1H NMR(500MHz,DMSO-d6)δ11.2(s,1H,NH),9.23(d,J=9.0Hz,1H,ArH),9.04(d,J=9.0Hz,1H,ArH),8.09(d,J=9.5Hz,1H,ArH),7.88(d,J=9.5Hz,1H,ArH),7.65(t,J=9.0Hz,1H,ArH),7.58(t,J=9.5Hz,1H,ArH),7.43(t,J=9.0Hz,2H,ArH),7.01(dd,J=10.0,8.5Hz,1H,H-1'),5.33(d,J=8.5Hz,1H,H-4'),4.31-4.36(m,1H,H-3'),2.93-2.98(m,1H,H-2'a),2.57(s,3H,N-CH3),2.04-2.11(m,1H,H-2'b),2.05(3H,s,6'-CH3);13C NMR(125MHz,DMSO-d6)δ170.8,170.5,155.5,141.3,137.6,129.8,128.0,127.0,126.5,124.8,124.5,123.5,121.2,121.0×2,120.8,119.8,117.4,116.5,115.7,109.3,92.6,79.0,75.3,52.0,29.6,28.2×2;ESIMS m/z 491.1[M-H]-.
化合物192,193,195和196的制备
将化合物191(10mg,0.020mmol)溶于10mL甲醇中,降至0℃加入硼氢化钠(7.6mg,0.2mmol),升至室温反应两小时,溶液由黄色变为无色,用乙酸乙酯稀释后加入饱和氯化铵溶液,乙酸乙酯萃取,无水硫酸钠干燥后蒸干。将粗产物溶于5mL冰醋酸中,加入锌粉(15mg,0.23mmol),升至40℃反应1.5小时,降至室温后,用乙酸乙酯稀释,饱和碳酸氢钠溶液洗涤后,无水硫酸钠干燥后浓缩。半制备HPLC分离、乙腈:水=7:13(v/v)洗脱得到4′-O-去甲基-(4′-O,3′-N)羰基-1′,5′-双表十字孢碱(192)(3.2mg,产率为35%)、5-脱氧-7-氧亚基-4′-O-去甲基-(4′-O,3′-N)羰基-1′,5′-双表十字孢碱(193)(3.2mg,产率为35%)。以化合物194(10mg)为原料,以相同的方法制得5-脱氧-7-氧亚基-4′-O-去甲基-(4′-O,3′-N)羰基十字孢碱(195)(2.7mg,产率为28%)和2.7mg 4′-O-去甲基-(4′-O,3′-N)羰基十字孢碱(ACT-007,196),产率为28%。
192:[α]D 20-78.2°(c 0.05,MeOH);1H NMR(600MHz,DMSO-d6)δ9.19(d,J=7.8Hz,1H,ArH),8.65(s,1H,NH),8.10(d,J=7.6Hz,1H,ArH),7.97(d,J=8.3Hz,1H,ArH),7.91(d,J=8.3Hz,1H,ArH),7.57(t,J=7.8Hz,1H,ArH),7.48-7.51(m,1H,ArH),7.43(t,J=7.5Hz,1H,ArH),7.29(t,J=7.5Hz,1H,ArH),6.92(dd,J=9.8,6.8Hz,1H,H-1'),5.72(d,J=9.7Hz,1H,H-4'),4.94-5.03(m,2H,H-7),4.41-4.44(m,1H,H-3'),2.98(s,3H,N-CH3),2.71-2.76(m,1H,H-2'a),2.07-2.12(m,1H,H-2'b),1.79(s,3H,6'-CH3).13C NMR(150MHz,DMSO-d6)δ171.6,156.9,138.8,136.9,133.2,128.4,125.6×2,125.5,124.9,124.8,122.3,122.0,121.2,120.6,119.6,116.1,115.2,113.6,109.4,93.3,77.4,71.1,52.0,45.5,28.9,25.0,24.4;HR-ESIMS m/z 479.1710[M+H]+(calcd for C28H23N4O4 +479.1719).
193:[α]D 20-76.5°(c 0.05,MeOH);1H NMR(600MHz,DMSO-d6)δ9.49(d,J=7.8Hz,1H,ArH),8.61(s,1H,NH),8.09(d,J=7.8Hz,1H,ArH),8.03(d,J=8.2Hz,1H,ArH),7.84(d,J=8.4Hz,1H,ArH),7.52-7.55(m,2H,ArH),7.36(t,J=7.5Hz,2H,ArH),6.93(dd,J=9.7,7.0Hz,1H,H-1'),5.76(d,J=9.5Hz,1H,H-4'),4.94-5.02(m,2H,H-5),4.44-4.46(m,1H,H-3'),2.99(s,3H,N-CH3),2.78(ddd,J=10.1,6.9,2.4Hz,1H,H-2'a),2.14-2.18(m,1H,H-2'b),1.75(s,3H,6'-CH3).13C NMR(150MHz,DMSO-d6)δ171.9,156.9,138.9,137.0,134.5,127.3,126.4×2,126.2,125.7,125.5,125.2,122.0,120.5,120.4,119.6,117.2,114.5,113.0,109.9,93.3,77.4,71.1,52.0,45.1,28.9,25.1,24.1;HR-ESIMS m/z 479.1711[M+H]+(calcd for C28H23N4O4 +479.1719).
195:[α]D 20+66°(c 0.05,MeOH);1H NMR(500MHz,DMSO-d6)δ9.51(d,J=7.9Hz,1H,ArH),8.61(s,1H,NH),8.10(d,J=7.7Hz,1H,ArH),8.01(d,J=8.5Hz,1H,ArH),7.85(d,J=8.2Hz,1H,ArH),7.56(t,J=7.6Hz,1H,ArH),7.46(t,J=7.6Hz,1H,ArH),7.37(t,J=7.4Hz,1H,ArH),7.30(t,J=7.4Hz,1H,ArH),6.96(dd,J=9.6,6.2Hz,1H,H-1'),5.30(d,J=8.7Hz,1H,H-4'),4.94-5.02(m,2H,H-5),4.32-4.37(m,1H,H-3'),2.92-2.96(m,1H,H-2'a),2.59(s,3H,N-CH3),2.05-2.10(m,1H,H-2'b),1.99(s,3H,6'-CH3).13C NMR(125MHz,DMSO-d6)δ171.8,155.6,140.3,136.5,134.2,126.9,126.5,125.6,125.4,125.0×2,122.0,121.7,120.4,119.9,119.3,117.6,115.9,114.1,109.2,92.4,79.1,75.4,52.0,44.9,29.4,28.7,28.2;HR-ESIMS m/z 479.1727[M+H]+(calcd for C28H23N4O4 +479.1719).
196:[α]D 20+73.2°(c 0.05,MeOH);1H NMR(500MHz,DMSO-d6)δ9.23(d,J=7.9Hz,1H,ArH),8.66(s,1H,NH),8.06(d,J=8.5Hz,1H,ArH),8.03(d,J=7.7Hz,1H,ArH),7.79(d,J=8.3Hz,1H,ArH),7.51(t,J=8.3Hz,1H,ArH),7.53(t,J=7.7Hz,1H,ArH),7.38(t,J=7.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),6.96(dd,J=9.7,6.3Hz,1H,H-1'),5.31(d,J=8.7Hz,1H,H-4'),4.95-5.03(m,2H,H-7),4.33-4.36(m,1H,H-3'),2.89-2.94(m,1H,H-2'a),2.59(s,3H,N-CH3),2.03(s,3H,6'-CH3),1.97-2.03(m,1H,H-2'b);13C NMR(125MHz,DMSO-d6)δ171.5,155.6,140.3,136.4,132.9,128.6,125.7,125.4,124.9,124.6×2,122.4,121.1,120.9,120.2,119.5,116.5,115.8,115.4,108.6,92.4,79.1,75.4,52.0,45.4,29.5,28.6,28.2;HR-ESIMS m/z 479.1725[M+H]+(calcd for C28H23N4O4 479.1719).
【实施例2】活性测试
1、α-葡萄糖苷酶抑制活性测试
(1)测试方法
以对硝基苯基-α-D-吡喃葡萄糖苷(PNPG)为底物,测试了部分化合物的α-葡萄糖苷酶抑制活性。PNPG是麦芽糖的类似物,它经过α-葡萄糖苷酶作用后能够生成显黄色的对硝基苯酚,可直接用于分光光度计的检测。具体是:首先样品用PBS的磷酸钠缓冲溶液(pH6.8)溶解配成5个浓度;然后在96孔板每孔中加入10μL样品溶液、20μL的PBS溶液、20μL的浓度为2.5mM的葡萄糖苷溶液(在磷酸盐缓冲溶液中溶解),于37℃培养箱中培养5分钟。用0.01M的PBS溶液将α-葡萄糖苷酶稀释成0.2U/mL,每孔取10μL加入到上述试液中,在37℃培养箱中培养15min后,用酶标仪测定每孔405nm处的吸光值(OD值),按下式计算抑制率(%):[1–(OD样品/OD对照)]×100%,求出IC50(μM)。空白对照为磷酸缓冲盐溶液、阳性对照为阿卡波糖和野尻霉素。
(2)实验结果
表1对α-葡萄糖苷酶抑制活性(IC50,μM)
结果表明,上述化合物对α-葡萄糖苷酶具有较强的抑制活性。
2、体内抗糖尿病肾病(DKD)实验
利用经典db/db糖尿病肾病动物模型评价了化合物49的体内抗糖尿病肾病(DKD)活性。
动物实验分6组,每组12只小鼠,阳性药物选择氯沙坦,各组均采用皮下注射给药的方式。连续给药4周,对照组给与相同体积的生理盐水。结果表明,在对空腹血糖(FBG)影响无明显变化的情况下,较低注射剂量的化合物49(1.0mg/kg/day和2.0mg/kg/day)能够显著降低24小时尿蛋白(24UPE)和血尿素氮(BUN)水平;特别是在1.0mg/kg/day的给药剂量时,对24UPE的抑制程度高达48.3%,优于阳性药物氯沙坦(Losartan)(35.2%)。结果见表2。
表2.化合物49的小鼠体内抗糖尿病肾病(DKD)活性
*p<0.05,**p<0.01vs.正常对照组;#p<0.05,##p<0.01vs.STZ对照组p;cp<0.05vs.氯沙坦组;a相对肾指数=左肾重量(g)/体重(g).数值为平均值±S.D.(n=12).
肾组织切片的H&E染色结果显示,糖尿病肾病db/db小鼠的肾小球系膜区明显扩张,细胞外基质增多,并出现弥散性系膜硬化,肾小球基底膜增厚;化合物49处理后显示出较好的治疗效果,特别是剂量在1.0mg/kg/day和2.0mg/kg/day时能够显著抑制肾组织的病理改变,减少系膜扩张,抑制弥散性系膜硬化;治疗效果优于阳性药物氯沙坦(图1)。